Benzamide and nicotinamide compounds and methods of using same

ABSTRACT

The present disclosure provides benzamide and nicotinamide compounds and pharmaceutical uses of the compounds. The compounds can be used to treat, for example, cancers such hematopoietic cancers (e.g., leukemia). The preferred compounds of the invention contain a phenylethynyl moiety as well as an amine-based heterocyclyl or heteroaryl moiety attached to the benzamide or nicotinamide compound.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to provisional patent application No.61/920,672 filed Dec. 24, 2014, the disclosure of which is herebyincorporated by reference.

BACKGROUND OF THE DISCLOSURE

Cancer continues to be a major health problem in the United States andworld-wide. In 2014, in the United States, there are expected to be morethan 1.5 million new cancer cases and more than 580,000 cancer deaths.Cancer-related deaths account for nearly ¼ of all deaths in the U.S.Most common childhood cancers are leukemias, lymphomas, brain tumors,and bone cancer, while adult cancers are more likely to be lung, colon,breast, prostate, and pancreas. Although enhanced early-stage tumordiagnosis and management have significantly increased patient survival,development and discovery of new anticancer therapies are still needed,in part because some patients exhibit insensitivity to currentanticancer drugs or develop drug-resistance after a period of treatment.

Leukemia is one of the most common hematologic malignancies in humanswhich usually begins in the bone marrow and results in high numbers ofabnormal white blood cells. Among acute leukemia, acute lymphoblasticleukemia (ALL) is a predominant cause of childhood leukemia, while acutemyeloid leukemia (AML) represents about 90% of all adult leukemia andthe second most common pediatric leukemia. While imatinib has improvedtherapy of chronic myelogenic leukemia due to specificity to its targetthe bcr-abl fusion gene product, the current treatment of ALL and AMLincludes cells proliferation affecting drugs that are not selective forhematologic malignancies, such as vincristine, anthracycline,cyclophosphamide etc. Such treatments often lead to severe side effects,development of resistance, and low survival rates.

BRIEF SUMMARY OF THE DISCLOSURE

In an aspect, the present disclosure provides benzamide and nicotinamidecompounds. The compounds can be used to selectively kill cancer cells(e.g., blood cancers). The compounds can be present

In various embodiments, the compounds have the following structure:

In these various embodiments, X is a carbon atom or nitrogen atom, Y isa single or triple bond, R¹ is selected from the group consisting of ahydrogen atom, a substituted or unsubstituted five to eight memberedheterocyclic ring, six membered aryl ring, five or six memberedheteroaryl ring, C₃ to C₈ cycloalkyl group, C₁ to C₆ alkyl group,

R² is selected from the group consisting of a hydrogen atom, asubstituted or unsubstituted five or six membered heteroaryl ring, afive or six membered aryl ring, C₃ to C₆ cycloalkyl group, eight to tenmembered heterocyclic ring system, and

R³ is selected from the group consisting of a substituted orunsubstituted C₂ to C₈ alkylheteroaryl group, C₂ to C₈alkyleneheteroaryl group, C₆ to C₁₀ aryl group, C₂ to C₅ heteroarylgroup, C₇ to C₁₃ alkylaryl group, C₇ to C₁₃ alkylenearyl group, C₂ to C₈alkylhetrocyclyl group, C₂ to C₈ alkylenehetrocyclyl group, C₄ to C₈alkylcycloalkyl group, C₄ to C₈ alkylenecycloalkyl group, or takentogether with R⁴ and the nitrogen atom to which they are attached form afive to seven membered substituted or unsubstituted heterocyclic ring.R⁴ is selected from the group consisting of a hydrogen atom andsubstituted or unsubstituted C₁ to C₆ alkyl group. R^(a) is asubstituted or unsubstituted C₁ to C₆ alkyl group or C₅ to C₆ arylgroup.

In an aspect, the present invention provides methods of using thecompounds. The compounds can be used, for example, to treat cancer.

In an embodiment, a method of treating cancer in an individual diagnosedwith or suspected of having cancer comprises administering to theindividual a therapeutically effective amount of one or more of thecompounds. In an embodiment, the cancer is a hematopoietic cancer. Thehematopoietic cancer is, for example, leukemia.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Example of growth of tumors in MV4-11 xenograft model of AML inSCID mice treated with vehicle control and TT-03197 at 10 mg/kg and 40mg/kg administered intraperitoneally. Mice were treated 6 days per weekas indicated on the figure. Results are Mean±SE (n=14-16).

FIG. 2. Example of growth of tumors in MV4-11 xenograft model of AML inSCID mice treated with vehicle control and TT-03203 at 10 mg/kg and 25mg/kg administered orally. Mice were treated 6 days per weeks asindicated on the figure. Results are Mean±SE (n=17-20)

FIG. 3. Example of survival of SCID mice inoculated via IV route withMV4-11 cells and treated via oral gavage with vehicle control, TT-01901at 100 mg/kg and TT-03586 at 50 mg/kg 6 days per week on days 4-58. Micewere sacrificed according to IACUC regulations after losing more than20% of body weight or becoming moribund and paralyzed.

DETAILED DESCRIPTION OF THE DISCLOSURE

In an aspect, the present disclosure provides benzamide and nicotinamidecompounds. The compounds can be used to selectively kill cancer cells(e.g., blood cancers).

In an embodiment, the present disclosure provides compounds having thefollowing structure (I):

where X is a carbon atom or nitrogen atom, Y is a single or triple bond,R¹ is selected from the group consisting of a hydrogen atom, asubstituted or unsubstituted five to eight membered heterocyclic ring,six membered aryl ring, five or six membered heteroaryl ring, C₃ to C₈cycloalkyl group, C₁ to C₆ alkyl group,

R² is selected from the group consisting of a hydrogen atom, asubstituted or unsubstituted five or six membered heteroaryl ring, afive or six membered aryl ring, C₃ to C₆ cycloalkyl group, eight to tenmembered heterocyclic ring system, and

R³ is selected from the group consisting of a substituted orunsubstituted C₂ to C₈ alkylheteroaryl group, C₂ to C₈alkyleneheteroaryl group, C₆ to C₁₀ aryl group, C₂ to C₅ heteroarylgroup, C₇ to C₁₃ alkylaryl group, C₇ to C₁₃ alkylenearyl group, C₂ to C₈alkylhetrocyclyl group, C₂ to C₈ alkylenehetrocyclyl group, C₄ to C₈alkylcycloalkyl group, C₄ to C₈ alkylenecycloalkyl group, or takentogether with R⁴ and the nitrogen atom to which they are attached form afive to seven membered substituted or unsubstituted heterocyclic ring;R⁴ is selected from the group consisting of a hydrogen atom andsubstituted or unsubstituted C₁ to C₆ alkyl group; and R^(a) is asubstituted or unsubstituted C₁ to C₆ alkyl group or C₅ to C₆ arylgroup.

As used herein, the term “alkyl group,” unless otherwise stated, refersto branched or unbranched hydrocarbons. Examples of such alkyl groupsinclude methyl groups, ethyl groups, propyl groups, butyl groups,isopropyl groups, tert-butyl groups, and the like. For example, thealkyl group can be a C₁ to C₆ alkyl group including all integer numbersof carbons and ranges of numbers of carbons therebetween. The alkylgroup can be unsubstituted or substituted with various substituents(e.g., as described herein).

As used herein, the term “alkylene,” unless otherwise stated refers toan alkyl group containing one or more double bonds.

As used herein, the term “aryl group,” unless otherwise stated refers toan aromatic carbocyclic group of 6 carbon atoms having a single ring(e.g., phenyl). The aryl group is substituted with 0, 1, 2, 3, 4, or 5substituents. The aryl group can be unsubstituted or substituted withvarious substituents (e.g., as described herein).

As used herein, the term “heteroaryl group,” unless otherwise statedrefers to an aromatic cyclic ring (i.e., fully unsaturated) having 1, 2,3, or 4 carbon atoms and 1, 2, 3, or 4 heteroatoms selected from oxygen,nitrogen, and sulfur. Examples of heteroaryl groups include thiophene,furan, and pyridine. The heteroaryl group is substituted with 0, 1, 2,3, or substituents. The heteroaryl group can be unsubstituted orsubstituted with various substituents as described herein.

As used herein, the term “cycloalkyl group,” unless otherwise stated,refers to a to a saturated or partially unsaturated carbocyclic group(not aromatic) of from 3 carbons to 6 carbons having a single cyclicring or multiple condensed rings. For example, the cycloalkyl groups canbe cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclohexene,cycloheptane, cycloheptene, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane,bicyclo[2.2.2]octane, bicyclo[3.3.0]octane, bicyclo[4.4.0]octane, andthe like. Cycloalkyl also includes carbocyclic groups to which is fusedan aryl or heteroaryl ring, for example indane andtetrahydronaphthalene. The cycloalkyl group can be unsubstituted orsubstituted with various substituents (e.g., as described herein).

As used herein, the term “heterocycle” or “heterocyclic ring,” unlessotherwise stated, refers to a cyclic compound having a ring where atleast one or more of the atoms forming the ring is a heteroatom (e.g.,oxygen, nitrogen, sulfur, etc.). The heterocyclic ring can be aromaticor nonaromatic, and include compounds that are saturated, partiallyunsaturated, and fully unsaturated. Examples of such groups includeazetidine, pyrrolidine, piperdine, azepane, azocane, dihydropyrdinone,dihydropyridazinone, dihydrooxepinone, dihydroazepinone, pyrazolone,pyrrolone, isoxazolone, pyranone, dihydrodiazepineone, furan, thiophene,oxazole, isoxazole, thiazole, oxadiazole, thiadiazole, triazole,tetrazole, oxazoline, lactam, lactone, dihydrofuran, tetrahydrofuran,furanone, oxazolone, pyridinone, pyrimidinone, dihydropyridazine,pyranone, oxazinone, and the like. For example, the heterocyclic ringcan be a 3, 4, 5, 6, 7, 8, 9 or 10 membered ring containing a number ofcarbon atoms ranging between 1 and 7 and a number of heteroatoms rangingbetween 1 and 7. The ring can be bonded to other rings to form ringsystems. The heterocyclic ring can be unsubstituted or substituted withvarious substituents (e.g., as described herein).

As used herein, the term “alkylheteroaryl group,” unless otherwisestated refers to an alkyl group, as defined herein, linked to aheteroaryl group as defined herein.

As used herein, the term “alkyleneheteroaryl group,” unless otherwisestated refers to an alkylene group, as defined herein, linked to aheteroaryl group as defined herein.

As used herein, the term “alkylaryl group,” unless otherwise statedrefers to an alkyl group, as defined herein, linked to a aryl group asdefined herein.

As used herein, the term “alkylenearyl group,” unless otherwise statedrefers to an alkylene group, as defined herein, linked to a aryl groupas defined herein.

As used herein, the term “alkylhetrocyclyl group,” unless otherwisestated refers to an alkyl group, as defined herein, linked to aheterocyclic ring as defined herein.

As used herein, the term “alkylenehetrocyclyl group,” unless otherwisestated refers to an alkylene group, as defined herein, linked to aheterocyclic ring as defined herein.

As used herein, the term “alkylcycloalkyl group,” unless otherwisestated refers to an alkyl group, as defined herein, linked to acycloalkyl group as defined herein.

As used herein, the term “alkylenecycloalkyl group,” unless otherwisestated refers to an alkylene group, as defined herein, linked to acycloalkyl group as defined herein.

As used herein, the term “substituents,” unless otherwise stated referto one or more of the following groups: alkyl groups, amines, alcoholgroups, alkoxy groups, halogen atoms, alkylhalides, alkylheteroarylgroups, alkoxy groups, hydroxyl groups, alkylalcohols, alkyl ethers,alkylamides, alkylamines, ketones, carbamates, PEG (polyethylene glycol)groups, cycloalkyl groups, alkyl esters, heteroaryl groups, aryl groups,nitriles, azido groups, amides, alkyenyl groups, alkynyl groups, thiolgroups, heterocyclyl groups, alkyleneheteroaryl groups, alkylarylgroups, alkylenearyl groups, alkylhetrocyclyl groups,alkylenehetrocyclyl groups, alkylcycloalkyl groups, andalkylenecycloalkyl groups.

In an embodiment, the disclosure provides compounds having the followingstructure (II):

where X, R¹, R², R³, and R⁴ are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructure (III):

where X, R¹, R², and R³ are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructure (IV):

where R⁵ is a C₂ to C₅ heteroaryl group and X, R¹, and R² are as definedherein.

In an embodiment, the disclosure provides compounds having the followingstructure (V):

where X, R¹, and R³ are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructure (VI):

where X, R², R³, and R⁴ are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructures (VII) and (VIII):

where X, R¹, and R² are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructures (IX) and (X):

where X and R¹ are as defined herein.

In an embodiment, the disclosure provides compounds having the followingstructures (XI) and (XII):

where X and R² are as defined herein.

In certain embodiments, R¹ is selected from the following groups:

where Z is N(R⁶)₂ or OR⁶, where each R⁶ is independently a hydrogen atomor substituted or unsubstituted C₁ to C₆ alkyl group.

In certain embodiments, R² is selected from the following groups:

where each R⁶ is independently a hydrogen atom or substituted orunsubstituted C₁ to C₆ alkyl group and n is 1, 2, 3, or 4.

In certain embodiments, the ring formed by R³—N—R⁴ is selected from thefollowing structures:

In certain embodiments, R³ is selected from the following groups:

In an embodiment, R⁴ is a hydrogen atom or a methyl group.

In an embodiment, R¹ is a substituted or unsubstituted five to eightmembered heterocyclic ring. For example, the five to eight memberedheterocyclic ring comprises at least one nitrogen atom.

In various embodiments, the compound of the present disclosure isselected from the following structures:

Structure StructureID

TT-01901

TT-01902

TT-02683

TT-02684

TT-02686

TT-02689

TT-02690

TT-02691

TT-02692

TT-02694

TT-02695

TT-02707

TT-02709

TT-02713

TT-02715

TT-02717

TT-02721

TT-02731

TT-02732

TT-02741

TT-02745

TT-02746

TT-02747

TT-02749

TT-02750

TT-02751

TT-02752

TT-02760

TT-02793

TT-02796

TT-02797

TT-02800

TT-02801

TT-02802

TT-02803

TT-02804

TT-02805

TT-02927

TT-02928

TT-02929

TT-02930

TT-02931

TT-02932

TT-02933

TT-02935

TT-02936

TT-02937

TT-02938

TT-02939

TT-02940

TT-02941

TT-02942

TT-02943

TT-02944

TT-02945

TT-02946

TT-02947

TT-02948

TT-02949

TT-03071

TT-03073

TT-03196

TT-03197

TT-03198

TT-03201

TT-03203

TT-03211

TT-03217

TT-03221

TT-03225

TT-03230

TT-03232

TT-03233

TT-03237

TT-03242

TT-03245

TT-03246

TT-03248

TT-03252

TT-03256

TT-03261

TT-03264

TT-03303

TT-03304

TT-03305

TT-03306

TT-03308

TT-03309

TT-03311

TT-03312

TT-03321

TT-03322

TT-03323

TT-03324

TT-03326

TT-03327

TT-03328

TT-03330

TT-03331

TT-03332

TT-03334

TT-03337

TT-03346

TT-03351

TT-03354

TT-03355

TT-03357

TT-03359

TT-03364

TT-03569

TT-03574

TT-03582

TT-03585

TT-03586

TT-03587

TT-03588

TT-03589

TT-03590

TT-03591

TT-03592

TT-03594

TT-03595

TT-03596

TT-03597

TT-03598

TT-03599

TT-03602

TT-03611

TT-03620

TT-03623

TT-03625

TT-03626

TT-03627

TT-03630

TT-03631

TT-03633

TT-03634

TT-03655

TT-03669

TT-03670

TT-03671

TT-03676

TT-03717

TT-03718

TT-03720

TT-03725

TT-03727

TT-03732

TT-03733

TT-03749

TT-03750

TT-03751

TT-03752

TT-03753

TT-03754

TT-03756

TT-03761

TT-03762

TT-03765

TT-03767

TT-03768

TT-03772

TT-03773

TT-03774

TT-03782

TT-03783

In an embodiment, the compound of the disclosure has the followingstructure:

In an embodiment, the compound of the disclosure is notN-(3-(1H-imidazol-1-yl)propyl)-3-(phenylethynyl)-4-(1H-pyrazol-1-yl)benzamide:

Non-limiting examples of general methods for the preparation of thecompounds of the present disclosure are provided in the followingschemes (i) and (ii):

More specific, non-limiting, examples of methods to synthesize compoundsof the present are illustrated in the examples that follow.

In an aspect, the present disclosure provides a composition comprisingat least one compound of the disclosure. Compositions comprising atleast one compound of the disclosure include, for example,pharmaceutical preparations.

The present disclosure includes all possible stereoisomers and geometricisomers of the benzamide or nicotinamide compounds (e.g., a compoundhaving the structure (I) to (XII)). The present disclosure includes bothracemic compounds and optically active isomers. When the benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)) is desired as a single enantiomer, it can be obtained either byresolution of the final product or by stereospecific synthesis fromeither isomerically pure starting material or use of a chiral auxiliaryreagent, for example, see Z. Ma et al., Tetrahedron: Asymmetry, 8(6),pages 883-888 (1997). Resolution of the final product, an intermediate,or a starting material can be achieved by any suitable method known inthe art. Additionally, in situations where tautomers of benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)) are possible, the present disclosure is intended to include alltautomeric forms of the compounds.

Prodrugs of the benzamide or nicotinamide compounds (e.g., a compoundhaving the structure (I) to (XII)) also can be used as the compound in amethod of the present disclosure. It is well established that a prodrugapproach, wherein a compound is derivatized into a form suitable forformulation and/or administration, then released as a drug in vivo, hasbeen successfully employed to transiently (e.g., bioreversibly) alterthe physicochemical properties of the compound (see, H. Bundgaard, Ed.,“Design of Prodrugs,” Elsevier, Amsterdam, (1985); R. B. Silverman, “TheOrganic Chemistry of Drug Design and Drug Action,” Academic Press, SanDiego, chapter 8, (1992); K. M. Hillgren et al., Med. Res. Rev., 15, 83(1995)).

Compounds of the present disclosure can contain one or more functionalgroups. The functional groups, if desired or necessary, can be modifiedto provide a prodrug. Suitable prodrugs include, for example, acidderivatives, such as amides and esters. It also is appreciated by thoseskilled in the art that N-oxides can be used as a prodrug.

Compounds of the disclosure can be in the form of salts.Pharmaceutically acceptable salts of the compounds of the disclosuregenerally are preferred in the methods of the disclosure. As usedherein, the term “pharmaceutically acceptable salts” refers to salts orzwitterionic forms of a benzamide or nicotinamide compound (e.g., acompound having the structure (I) to (XII)). Salts of benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)) can be prepared during the final isolation and purification ofthe compounds or separately by reacting the compound with an acid havinga suitable cation. In an embodiment, the pharmaceutically acceptablesalts of a benzamide or nicotinamide compounds (e.g., a compound havingthe structure (I) to (XII)) are acid addition salts formed withpharmaceutically acceptable acids. Examples of acids which can beemployed to form pharmaceutically acceptable salts include inorganicacids such as nitric, boric, hydrochloric, hydrobromic, sulfuric, andphosphoric, and organic acids such as oxalic, maleic, succinic, andcitric. Nonlimiting examples of salts of compounds of the disclosureinclude, but are not limited to, the hydrochloride, hydrobromide,hydroiodide, sulfate, bisulfate, 2-hydroxyethansulfonate, phosphate,hydrogen phosphate, acetate, adipate, alginate, aspartate, benzoate,bisulfate, butyrate, camphorate, camphorsulfonate, digluconate,glycerolphsphate, hemisulfate, heptanoate, hexanoate, formate,succinate, fumarate, maleate, ascorbate, isethionate, salicylate,methanesulfonate, mesitylenesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate,3-phenylproprionate, picrate, pivalate, propionate, trichloroacetate,trifluoroacetate, phosphate, glutamate, bicarbonate,paratoluenesulfonate, undecanoate, lactate, citrate, tartrate,gluconate, methanesulfonate, ethanedisulfonate, benzene sulphonate, andp-toluenesulfonate salts. In addition, available amino groups present inthe compounds of the disclosure can be quaternized with methyl, ethyl,propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl,dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and sterylchlorides, bromides, and iodides; and benzyl and phenethyl bromides. Inlight of the foregoing, any reference to compounds of the presentdisclosure appearing herein is intended to include benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)) as well as pharmaceutically acceptable salts, hydrates, orprodrugs thereof.

A benzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) also can be conjugated or linked to auxiliarymoieties that promote a beneficial property of the compound in a methodof therapeutic use. Such conjugates can enhance delivery of thecompounds to a particular anatomical site or region of interest (e.g., atumor), enable sustained therapeutic concentrations of the compounds intarget cells, alter pharmacokinetic and pharmacodynamics properties ofthe compounds, and/or improve the therapeutic index or safety profile ofthe compounds. Suitable auxiliary moieties include, for example, aminoacids, oligopeptides, or polypeptides, e.g., antibodies such asmonoclonal antibodies and other engineered antibodies; and natural orsynthetic ligands to receptors in target cells or tissues. Othersuitable auxiliaries include fatty acid or lipid moieties that promotebiodistribution and/or uptake of the compound by target cells (see,e.g., Bradley et al., Clin. Cancer Res. (2001) 7:3229).

In an aspect, the disclosure provides a method of treating cancer in anindividual diagnosed with or suspected of having cancer comprisingadministering to the individual a therapeutically effective amount of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) as described herein. In an embodiment, thecancer is a blood cancer. The blood cancer can be, for example,leukemia.

The language “therapeutically effective amount” of a compound of thedisclosure refers to an amount of an agent which is effective, uponsingle or multiple dose administration to the patient, in inhibitingcell proliferation and/or symptoms of a cell proliferative disorder, orin prolonging the survivability of the patient with such a cellproliferative disorder beyond that expected in the absence of suchtreatment. The exact amount desired or required may vary depending onthe particular compound or composition used, its mode of administration,and the like. Appropriate effective amount can be determined by one ofordinary skill in the art informed by the instant disclosure using onlyroutine experimentation.

Within the meaning of the disclosure, “treatment” also includes relapseprophylaxis or phase prophylaxis, as well as the treatment of acute orchronic signs, symptoms and/or malfunctions. The treatment can beorientated symptomatically, for example, to suppress symptoms. It can beeffected over a short period, be oriented over a medium term, or can bea long-term treatment, for example within the context of a maintenancetherapy.

Compositions comprising a compound of the disclosure and apharmaceutical agent can be prepared at a patient's bedside, or by apharmaceutical manufacture. In the latter case, the compositions can beprovided in any suitable container, such as a sealed sterile vial orampoule, and may be further packaged to include instruction documentsfor use by a pharmacist, physician or other health care provider. Thecompositions can be provided as a liquid, or as a lyophilized or powderform that can be reconstituted if necessary when ready for use. Thecompositions can be provided in combination with any suitable deliveryform or vehicle, examples of which include, for example, liquids,caplets, capsules, tablets, inhalants or aerosol, etc. The deliverydevices may comprise components that facilitate release of thepharmaceutical agents over certain time periods and/or intervals, andcan include compositions that enhance delivery of the pharmaceuticals,such as nanoparticle, microsphere or liposome formulations, a variety ofwhich are known in the art and are commercially available. Further, eachcomposition described herein can comprise one or more pharmaceuticalagents. The compositions described herein can include one or morestandard pharmaceutically acceptable carriers. Examples ofpharmaceutically acceptable carriers can be found in: Remington: TheScience and Practice of Pharmacy (2005) 21st Edition, Philadelphia, Pa.Lippincott Williams & Wilkins.

Various methods known to those skilled in the art can be used tointroduce the compositions of the disclosure to an individual. Thesemethods, for example, of introducing the benzamide or nicotinamidecompound, or compositions containing the benzamide or nicotinamidecompound, can be administered in any manner including, but not limitedto, orally, parenterally, sublingually, transdermally, rectally,transmucosally, topically, via inhalation, via buccal administration, orcombinations thereof. Parenteral administration includes, but is notlimited to, intravenous, intra-arterial, intracranial, intradermal,subcutaneous, intraperitoneal, subcutaneous, intramuscular, intrathecal,and intraarticular. The benzamide or nicotinamide compound also can beadministered in the form of an implant, which allows a slow release ofthe compound, as well as a slow controlled i.v. infusion.

The dose of the composition comprising a compound of the disclosure anda pharmaceutical agent generally depends upon the needs of theindividual to whom the composition of the disclosure is to beadministered. These factors include, for example, the weight, age, sex,medical history, and nature and stage of the disease for which atherapeutic or prophylactic effect is desired. The compositions can beused in conjunction with any other conventional treatment modalitydesigned to improve the disorder for which a desired therapeutic orprophylactic effect is intended, non-limiting examples of which includesurgical interventions and radiation therapies. The compositions can beadministered once, or over a series of administrations at variousintervals determined using ordinary skill in the art, and given thebenefit of the present disclosure.

Compositions of the disclosure can comprise more than one pharmaceuticalagent. For example, a first composition comprising a compound of thedisclosure and a first pharmaceutical agent can be separately preparedfrom a composition which comprises the same compound of the disclosureand a second pharmaceutical agent, and such preparations can be mixed toprovide a two-pronged (or more) approach to achieving the desiredprophylaxis or therapy in an individual. Further, compositions of thedisclosure can be prepared using mixed preparations of any of thecompounds disclosed herein.

It is envisioned, therefore, that a benzamide or nicotinamide compound(e.g., a compound having the structure (I) to (XII)) are useful in thetreatment of a variety of conditions and diseases. Thus, the presentdisclosure concerns the use of benzamide or nicotinamide compounds(e.g., a compound having the structure (I) to (XII)), or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition containing either entity, for the manufacture of amedicament for the treatment of such conditions and diseases.

The compounds of the present disclosure can be therapeuticallyadministered as the neat chemical, but it is preferred to administer abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) as a pharmaceutical composition or formulation.Thus, the present disclosure provides a pharmaceutical compositioncomprising a benzamide or nicotinamide compound (e.g., a compound havingthe structure (I) to (XII)) together with a pharmaceutically acceptablediluent or carrier therefor. Also provided is a process of preparing apharmaceutical composition comprising admixing a benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) with a pharmaceutically acceptable diluent or carrier therefor.

Accordingly, the present disclosure further provides pharmaceuticalformulations comprising a benzamide or nicotinamide compound (e.g., acompound having the structure (I) to (XII)), or a pharmaceuticallyacceptable salt, prodrug, or hydrate thereof, together with one or morepharmaceutically acceptable carriers and, optionally, other therapeuticand/or prophylactic ingredients. The carriers are “acceptable” in thesense of being compatible with the other ingredients of the formulationand not deleterious to the recipient thereof.

Examples of pharmaceutically-acceptable carrier includepharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting the subject chemical fromone organ, or portion of the body, to another organ, or portion of thebody. Some examples of materials which can serve aspharmaceutically-acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically-acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

In an embodiment, the pharmaceutically-acceptable formulation is suchthat it provides sustained delivery of a benzamide or nicotinamidecompound (e.g., a compound having the structure (I) to (XII)) to asubject for at least 3 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48hours, one week, two weeks, three weeks, or four weeks after thepharmaceutically-acceptable formulation is administered to the subject.

In certain embodiments, these pharmaceutical compositions are suitablefor topical or oral administration to a subject. In other embodiments,as described in detail below, the pharmaceutical compositions of thepresent disclosure may be specially formulated for administration insolid or liquid form, including those adapted for the following: (1)oral administration, for example, drenches (aqueous or non-aqueoussolutions or suspensions), tablets, boluses, powders, granules, pastes;(2) parenteral administration, for example, by subcutaneous,intramuscular or intravenous injection as, for example, a sterilesolution or suspension; (3) topical application, for example, as acream, ointment or spray applied to the skin; (4) intravaginally orintrarectally, for example, as a pessary, cream or foam; or (5) aerosol,for example, as an aqueous aerosol, liposomal preparation or solidparticles.

The compositions may conveniently be presented in unit dosage form andmay be prepared by any methods well known in the art of pharmacy. Theamount of active ingredient which can be combined with a carriermaterial to produce a single dosage form will vary depending upon thehost being treated, the particular mode of administration. The amount ofactive ingredient which can be combined with a carrier material toproduce a single dosage form will generally be that amount of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) which produces a therapeutic effect. Generally,out of one hundred percent, this amount will range from about 1 percentto about ninety-nine percent of active ingredient, preferably from about5 percent to about 70 percent, more preferably from about 10 percent toabout 30 percent.

Methods of preparing these compositions include the step of bringinginto association a benzamide or nicotinamide compound (e.g., a compoundhaving the structure (I) to (XII)) with the carrier and, optionally, oneor more accessory ingredients. In general, the formulations are preparedby uniformly and intimately bringing into association a benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) with liquid carriers, or finely divided solid carriers, or both,and then, if necessary, shaping the product.

Compositions of the disclosure suitable for oral administration may bein the form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) as an active ingredient. A benzamide or nicotinamide compound(e.g., a compound having the structure (I) to (XII)) may also beadministered as a bolus, electuary or paste.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered activeingredient moistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present disclosure, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active ingredient can also be in micro-encapsulated form,if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of a benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) include pharmaceutically-acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof

In addition to inert diluents, the oral compositions can includeadjuvants such as wetting agents, emulsifying and suspending agents,sweetening, flavoring, coloring, perfuming and preservative agents.

Pharmaceutical compositions of the disclosure for rectal or vaginaladministration may be presented as a suppository, which may be preparedby mixing a benzamide or nicotinamide compound (e.g., a compound havingthe structure (I) to (XII)) with one or more suitable nonirritatingexcipients or carriers comprising, for example, cocoa butter,polyethylene glycol, a suppository wax or a salicylate, and which issolid at room temperature, but liquid at body temperature and,therefore, will melt in the rectum or vaginal cavity and release theactive agent.

Dosage forms for the topical or transdermal administration of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) include powders, sprays, ointments, pastes,creams, lotions, gels, solutions, patches and inhalants. A benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) may be mixed under sterile conditions with apharmaceutically-acceptable carrier, and with any preservatives,buffers, or propellants which may be required.

The ointments, pastes, creams and gels may contain, in addition to abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)), excipients, such as animal and vegetable fats,oils, waxes, paraffins, starch, tragacanth, cellulose derivatives,polyethylene glycols, silicones, bentonites, silicic acid, talc and zincoxide, or mixtures thereof

Powders and sprays can contain, in addition to a benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)), excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Ordinarily, an aqueous aerosol is made by formulating an aqueoussolution or suspension of the agent together with conventionalpharmaceutically-acceptable carriers and stabilizers. The carriers andstabilizers vary with the requirements of the particular benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)), but typically include nonionic surfactants (Tweens, Pluronics,or polyethylene glycol), innocuous proteins like serum albumin, sorbitanesters, oleic acid, lecithin, amino acids such as glycine, buffers,salts, sugars or sugar alcohols. Aerosols generally are prepared fromisotonic solutions.

Transdermal patches have the added advantage of providing controlleddelivery of a benzamide or nicotinamide compound (e.g., a compoundhaving the structure (I) to (XII)) to the body. Such dosage forms can bemade by dissolving or dispersing the agent in the proper medium.Absorption enhancers can also be used to increase the flux of the activeingredient across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the activeingredient in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of the disclosure.

Pharmaceutical compositions of the disclosure suitable for parenteraladministration comprise a benzamide or nicotinamide compound (e.g., acompound having the structure (I) to (XII)) in combination with one ormore pharmaceutically-acceptable sterile isotonic aqueous or nonaqueoussolutions, dispersions, suspensions or emulsions, or sterile powderswhich may be reconstituted into sterile injectable solutions ordispersions just prior to use, which may contain antioxidants, buffers,bacteriostats, solutes which render the formulation isotonic with theblood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers, which may beemployed in the pharmaceutical compositions of the disclosure includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents which delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue.

When a benzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) are administered as pharmaceuticals, toindividuals (e.g., to humans and non-humans (i.e., animals)), they canbe given per se or as a pharmaceutical composition containing, forexample, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredientin combination with a pharmaceutically-acceptable carrier.

Regardless of the route of administration selected, a compound havingthe structure (I) to (XII), which may be used in a suitable hydratedform, and/or the pharmaceutical compositions of the present disclosure,are formulated into pharmaceutically-acceptable dosage forms byconventional methods known to those of skill in the art.

In certain embodiments, the methods of the disclosure includeadministering to a subject a therapeutically effective amount of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) in combination with another pharmaceuticallyactive ingredient. Examples of pharmaceutically active ingredients knownto treat cell proliferative disorders, e.g., anticancer agent,antiproliferative agent, chemotherapeutic. Other pharmaceutically activeingredients that may be used can be found in Harrison's Principles ofInternal Medicine, Thirteenth Edition, Eds. T. R. Harrison et al.McGraw-Hill N.Y., NY; and the Physicians Desk Reference 50th Edition1997, Oradell New Jersey, Medical Economics Co., the complete contentsof which are expressly incorporated herein by reference. A benzamide ornicotinamide compound (e.g., a compound having the structure (I) to(XII)) and the pharmaceutically active ingredient may be administered tothe subject in the same pharmaceutical composition or in differentpharmaceutical compositions (at the same time or at different times).

Methods delineated herein include those wherein the subject isidentified as in need of a particular stated treatment. Identifying asubject in need of such treatment can be in the judgment of a subject ora health care professional and can be subjective (e.g. opinion) orobjective (e.g. measurable by a test or diagnostic method). In othermethods, the subject is prescreened or identified as in need of suchtreatment by assessment for a relevant marker or indicator ofsuitability for such treatment. The identification of those patients whoare in need of prophylactic treatment for cell proliferative disordersis well within the ability and knowledge of one skilled in the art.Certain of the methods for identification of patients which are at riskof developing cell proliferative disorders which can be treated by thesubject method are appreciated in the medical arts, such as familyhistory, and the presence of risk factors associated with thedevelopment of that disease state in the subject patient. A clinicianskilled in the art can readily identify such candidate patients, by theuse of, for example, clinical tests, physical examination andmedical/family history. The subject may have a cell proliferativedisorder, may be at risk of developing a cell proliferative disorder, ormay need prophylactic treatment prior to anticipated or unanticipatedexposure to a condition(s) capable of increasing susceptibility to acell proliferative disorder, e.g., exposure to carcinogens or toionizing radiation.

In an aspect, the disclosure provides a kit for treating a cellproliferative disorder in a subject is provided and includes a benzamideor nicotinamide compound (e.g., a compound having the structure (I) to(XII)), pharmaceutically acceptable esters, salts, and prodrugs thereof,and instructions for use. In further aspects, the disclosure provideskits for inhibiting cell proliferation, assessing the efficacy of ananti-cell proliferative treatment in a subject, monitoring the progressof a subject being treated with a cell proliferation inhibitor,selecting a subject with a cell proliferative disorder for treatmentwith cell proliferation inhibitor, and/or treating a subject sufferingfrom or susceptible to cancer. In certain embodiments, the disclosureprovides: a kit for treating a cell proliferative disorder in a subject,the kit comprising a benzamide or nicotinamide compound (e.g., acompound having the structure (I) to (XII)).

For veterinary use, a benzamide or nicotinamide compound (e.g., acompound having the structure (I) to (XII)), or a pharmaceuticallyacceptable salt or prodrug, is administered as a suitably acceptableformulation in accordance with normal veterinary practice. Theveterinarian can readily determine the dosing regimen and route ofadministration that is most appropriate for a particular animal. Animalstreatable by the present compounds and methods include, but are notlimited to, pets, livestock, show animals, and zoo specimens.

When administered in combination with other therapeutics, a presentbenzamide or nicotinamide compound may be administered at relativelylower dosages. In addition, the use of targeting agents may allow thenecessary dosage to be relatively low. Certain compounds may beadministered at relatively high dosages due to factors including, butnot limited to, low toxicity and high clearance.

For human use, a benzamide or nicotinamide compound (e.g., a compoundhaving the structure (I) to (XII)) can be administered alone, butgenerally is administered in admixture with a pharmaceutical carrierselected with regard to the intended route of administration andstandard pharmaceutical practice. Pharmaceutical compositions for use inaccordance with the present disclosure can be formulated in aconventional manner using one or more physiologically acceptable carriercomprising excipients and auxiliaries that facilitate processing of abenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) into pharmaceutical preparations.

The present benzamide or nicotinamide compounds can be administeredsimultaneously or metronomically with other anti-cancer treatments, suchas chemotherapy and/or radiation therapy. The term “simultaneous” or“simultaneously” means that the other anti-cancer treatment and thebenzamide or nicotinamide compound are administered within 6 hours, 3hours or less, of each other. The term “metronomically” means theadministration of the other anti-cancer treatments at times differentfrom the anti-cancer treatments and at a certain frequency relative torepeat administration and/or the anti-cancer treatment regimen.

The benzamide or nicotinamide compounds of the present disclosure can beused to treat a variety of diseases and conditions. For example,compounds of the present disclosure can be used in combination withradiation and/or a chemotherapeutic agent in the treatment of cancers.For example, the benzamide or nicotinamide compounds can be used toenhance treatment of tumors that are customarily treated with anantimetabolite, e.g., methotrexate or 5-fluorouracil (5-FU).

Use of benzamide or nicotinamide compounds of the present disclosure canresult in partial or complete regression of cancer cells, i.e., thepartial or complete disappearance of such cells from the cellpopulation. For example, a method of the disclosure can be used to slowthe rate of tumor growth, decrease the size or number of tumors, or toinduce partial or complete tumor regression.

In an embodiment, cancers treated by benzamide or nicotinamide compounds(e.g., a compound having the structure (I) to (XII)) are hematopoietictumors of lymphoid lineage, including leukemia, acute lymphocyticleukemia, acute lymphoblastic leukemia, 8-celllymphoma, T-cell lymphoma,Hodgkins lymphoma, nonHodgkins lymphoma, hairy cell lymphoma,histiocytic lymphoma, and Burketts lymphoma, hematopoietic tumors ofmyeloid lineage, including acute and chronic myelogenous leukemias,myelodysplastic syndrome, myeloid leukemia, and promyelocytic leukemia.

A benzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) can be used for treating a disease or conditionin vivo by administration to an individual in need thereof. The diseaseor condition can be a cancer. A variety of cancers can be treatedincluding, but not limited to: carcinomas, including bladder (includingaccelerated and metastic bladder cancer), breast, colon (includingcolorectal cancer), kidney, liver, lung (including small and non-smallcell lung cancer and lung adenocarcinoma), ovary, prostate, testes,genitourinary tract, lymphatic system, rectum, larynx, pancreas(including exocrine pancreatic carcinoma), esophagus, stomach, gallbladder, cervix, thyroid, renal, and skin (including squamous cellcarcinoma); tumors of the central and peripheral nervous system,including astrocytoma, neuroblastoma, glioma, and schwannomas; tumors ofmesenchymal origin, including fibrosarcoma, rhabdomyoscarcoma, andosteosarcoma; and other tumors, including melanoma, xenodermapigmentosum, keratoactanthoma, seminoma, thyroid follicular cancer,teratocarcinoma, renal cell carcinoma (RCC), pancreatic cancer, myeloma,myeloid and lymphoblastic leukemia, neuroblastoma, and glioblastoma.

One method of the present disclosure comprises administration of atherapeutically effective amount of a present benzamide or nicotinamidecompound in combination with a chemotherapeutic agent that can effectsingle- or double-strand DNA breaks or that can block DNA replication orcell proliferation. Alternatively, a method of the present disclosurecomprises administration of a therapeutically effective amount of atleast one present benzamide or nicotinamide compound in combination withtherapies that include use of an antibody, e.g., herceptin, that hasactivity in inhibiting the proliferation of cancer cells. Accordingly,cancers, for example, colorectal cancers, head and neck cancers,pancreatic cancers, breast cancers, gastric cancers, bladder cancers,vulvar cancers, leukemias, lymphomas, melanomas, renal cell carcinomas,ovarian cancers, brain tumors, osteosarcomas, and lung carcinomas, aresusceptible to enhanced treatment by administration of a presentbenzamide or nicotinamide in combination with a chemotherapeutic agentor an antibody.

Without intending to be bound by any particular theory it is consideredthat compounds of the present disclosure inhibit NAMPT (Nicotinamidephosphoribosyltransferase). It is considered that based on suchinhibition that compounds of the present disclosure have efficacyagainst diseases related to this target such as acute respiratorydistress syndrome (ARDS), aging, atherosclerosis, cancer, diabetesmellitus, rheumatoid arthritis, and sepsis.

Cancers treatable by the present disclosure also include solid tumors,i.e., carcinomas and sarcomas. Carcinomas include malignant neoplasmsderived from epithelial cells which infiltrate (i.e., invade)surrounding tissues and give rise to metastases. Adenocarcinomas arecarcinomas derived from glandular tissue, or from tissues that formrecognizable glandular structures. Another broad category of cancersincludes sarcomas, which are tumors whose cells are embedded in afibrillar or homogeneous substance, like embryonic connective tissue.The present disclosure also enables treatment of cancers of the myeloidor lymphoid systems, including leukemias, lymphomas, and other cancersthat typically are not present as a tumor mass, but are distributed inthe vascular or lymphoreticular systems.

Additional forms of cancer treatable by the present benzamide ornicotinamide compounds include, for example, adult and pediatriconcology, growth of solid tumors/malignancies, myxoid and round cellcarcinoma, locally advanced tumors, metastatic cancer, human soft tissuesarcomas, including Ewing's sarcoma, cancer metastases, includinglymphatic metastases, squamous cell carcinoma, particularly of the headand neck, esophageal squamous cell carcinoma, oral carcinoma, blood cellmalignancies, including multiple myeloma, leukemias, including acutelymphocytic leukemia, acute nonlymphocytic leukemia, chronic lymphocyticleukemia, chronic myelocytic leukemia, and hairy cell leukemia, effusionlymphomas (body cavity based lymphomas), thymic lymphoma lung cancer(including small cell carcinoma, cutaneous T cell lymphoma, Hodgkin'slymphoma, non-Hodgkin's lymphoma, cancer of the adrenal cortex,ACTH-producing tumors, nonsmall cell cancers, breast cancer, includingsmall cell carcinoma and ductal carcinoma), gastrointestinal cancers(including stomach cancer, colon cancer, colorectal cancer, and polypsassociated with colorectal neoplasia), pancreatic cancer, liver cancer,urological cancers (including bladder cancer, such as primarysuperficial bladder tumors, invasive transitional cell carcinoma of thebladder, and muscleinvasive bladder cancer), prostate cancer,malignancies of the female genital tract (including ovarian carcinoma,primary peritoneal epithelial neoplasms, cervical carcinoma, uterineendometrial cancers, vaginal cancer, cancer of the vulva, uterine cancerand solid tumors in the ovarian follicle), malignancies of the malegenital tract (including testicular cancer and penile cancer), kidneycancer (including renal cell carcinoma, brain cancer (includingintrinsic brain tumors, neuroblastoma, astrocytic brain tumors, gliomas,and metastatic tumor cell invasion in the central nervous system), bonecancers (including osteomas and osteosarcomas), skin cancers (includingmalignant melanoma, tumor progression of human skin keratinocytes, andsquamous cell cancer), thyroid cancer, retinoblastoma, neuroblastoma,peritoneal effusion, malignant pleural effusion, mesothelioma, Wilms'stumors, gall bladder cancer, trophoblastic neoplasms,hemangiopericytoma, and Kaposi's sarcoma. Accordingly, administration ofa present benzamide or nicotinamide compound is expected to enhancetreatment regimens.

In an aspect, a benzamide or nicotinamide compound (e.g., a compoundhaving the structure (I) to (XII)) exhibit anticancer activity. Invarious embodiments, the compounds of the disclosure are those in whichshow IC50 values in cytotoxicity experiments towards cells lines MV4-11and U937 of >20 μM, 10-20 μM, 5-10 μM, 1-5 μM, or <1 μM.

As appreciated by persons skilled in the art, additional active orancillary agents can be used in the methods described herein. Referenceherein to treatment also extends to prophylaxis, as well as to treatmentof established diseases or symptoms.

The compound of the present disclosure can be applied to cellpopulations ex vivo. For example, the present benzamide or nicotinamidecompounds can be used ex vivo to determine the optimal schedule and/ordosing of administration of the present benzamide or nicotinamidecompound for a given indication, cell type, patient, and otherparameter. Information gleaned from such use can be used forexperimental purposes or in the clinic to set protocol for in vivotreatment. Other ex vivo uses for which the disclosure is suited areapparent to those skilled in the art.

A present benzamide or nicotinamide compound also can be administered incombination with radiation. Diseases that are treatable withelectromagnetic radiation include neoplastic diseases, benign andmalignant tumors, and cancerous cells.

Electromagnetic radiation treatment of other diseases not listed hereinalso is contemplated by the present disclosure. Preferred embodiments ofthe present disclosure employ the electromagnetic radiation of:gamma-radiation (10-20 to 10-13 m), X-ray radiation (10-12 to 10-9 m),ultraviolet light (10 nm to 400 nm), visible light (400 nm to 700 nm),infrared radiation (700 nm to 1 mm), and microwave radiation (1 mm to 30nm).

Many cancer treatment protocols currently employ radiosensitizersactivated by electromagnetic radiation, e.g., X-rays. Examples ofX-ray-activated radiosensitizers include, but are not limited to, thefollowing: metronidazole, misonidazole, desmethylmisonidazole,pimonidazole, etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233,E09, RB 6145, nicotinamide, 5-bromodeoxyuridine (BUdR),5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine(FUdR), hydroxyurea, cis-platin, and therapeutically effective analogsand derivatives of the same.

Photodynamic therapy (PDT) of cancers employs visible light as theradiation activator of the sensitizing agent. Examples of photodynamicradiosensitizers include the following, but are not limited to:hematoporphyrin derivatives, PHOTOFRINriD, benzoporphyrin derivatives,NPe6, tin etioporphyrin (SnET2), pheoborbide-a, bacteriochlorophyll-a,naphthalocyanines, phthalocyanines, zinc phthalocyanine, andtherapeutically effective analogs and derivatives of the same.

Radiosensitizers can be administered in conjunction with atherapeutically effective amount of one or more compounds in addition toa present benzamide or nicotinamide compound, such compounds including,but not limited to, compounds that promote the incorporation ofradiosensitizers to the target cells, compounds that control the flow oftherapeutics, nutrients, and/or oxygen to the target cells,chemotherapeutic agents that act on the tumor with or without additionalradiation, or other therapeutically effective compounds for treatingcancer or other disease. Examples of additional therapeutic agents thatcan be used in conjunction with radiosensitizers include, but are notlimited to, 5-fluorouracil (5-FU), leucovorin, oxygen, carbogen, redcell transfusions, perfluorocarbons (e.g., FLUOSOLW®-DA), 2,3-DPG,BW12C, calcium channel blockers, pentoxifylline, antiangiogenesiscompounds, hydralazine, and L-BSO.

The chemotherapeutic agent can be any pharmacological agent or compoundthat induces apoptosis. The pharmacological agent or compound can be,for example, a small organic molecule, peptide, polypeptide, nucleicacid, or antibody. Chemotherapeutic agents that can be used include, butare not limited to, alkylating agents, antimetabolites, hormones andantagonists thereof, natural products and their derivatives,radioisotopes, antibodies, as well as natural products, and combinationsthereof. For example, a benzamide or nicotinamide compound of thepresent disclosure can be administered with antibiotics, such asdoxorubicin and other anthracycline analogs, nitrogen mustards, such ascyclophosphamide, pyrimidine analogs such as 5-fluorouracil, cis-platin,hydroxyurea, taxol and its natural and synthetic derivatives, and thelike. As another example, in the case of mixed tumors, such asadenocarcinoma of the breast, where the tumors includegonadotropin-dependent and gonadotropin-independent cells, the compoundcan be administered in conjunction with leuprolide or goserelin(synthetic peptide analogs of LH-RH). Other antineoplastic protocolsinclude the use of an inhibitor compound with another treatmentmodality, e.g., surgery or radiation, also referred to herein as“adjunct anti-neoplastic modalities.” Additional chemotherapeutic agentsuseful in the disclosure include hormones and antagonists thereof,radioisotopes, antibodies, natural products, and combinations thereof

A benzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)) of this disclosure can be provided inpharmaceutical compositions. In an embodiment, the pharmaceuticalcomposition comprises one or more benzamide or nicotinamide compounds(e.g., a compound having the structure (I) to (XII)) of the presentdisclosure and a pharmaceutically acceptable carrier. In an embodiment,the kits of the disclosure can comprise one or more benzamide ornicotinamide compounds (e.g., a compound having the structure (I) to(XII)) alone, as pharmaceutical preparations, or separate pharmaceuticalpreparations with each pharmaceutical preparation comprising a separatebenzamide or nicotinamide compound (e.g., a compound having thestructure (I) to (XII)).

The following specific examples are to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever.

Example 1

This example provides synthetic procedures for benzamides andnicotinamides of the present disclosure.

Procedure for Synthesis of Compound 24.

A suspension of compound 14 (33.6 g, 156 mmol, 1.0 eq.) and hydrazinehydrate (12 mL, 386 mmol, 2.5 eq.) in ethanol (280 mL) was stirred atroom temperature for 20 hours. The formed precipitate was filtered offand dried. The obtained product was purified by column chromatography(silica gel, hexane/ethyl acetate, 2:1) giving compound 24 (10.86 g,33%) as a yellow-orange solid. APCI-MS (m/z (intensity)): 211.70([M+H]⁺, 90%), 253.13 ([M+MeCN+H]⁺, 100%).

Procedure for Synthesis of Compound 25.

A mixture of concentrated aqueous HCl solution (0.75 mL, 8.6 mmol, 0.17eq.) and water (37 mL) was added to a suspension of compound 24 (10.85g, 51.38 mmol, 1.0 eq.) and 1,1,3,3-tetramethoxypropane (12.50 g, 76.12mmol, 1.5 eq.) in EtOH (74 mL) dropwise at room temperature. Thereaction mixture was stirred at refluxing for 2.5 hours. The formedprecipitate was collected by filtration and dried. The obtained productwas purified by column chromatography (silica gel, hexane/ethyl acetate,4:1) giving compound 25 (12.05 g, 95%) as a yellowish crystals. APCI-MS(m/z (intensity)): 248.10 ([M+H]⁺, 100%), 289.12 ([M+MeCN+H]⁺, 10%).

Procedure for Synthesis of Compound 26.

A mixture of compound 25 (12.05 g. 48.74 mmol, 1.0 eq.), Raney nickelcatalyst (2.4 g, 40.89 mmol, 0.84 eq.) and methanol (500 mL) washydrogenated (2 atm) at room temperature for 16 hours. The catalyst wasremoved by filtration. The filtrate was concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, hexane/ethyl acetate, 4:1) giving compound 26 (9.91 g, 94%)as a yellowish solid. APCI-MS (m/z (intensity)): 218.10 ([M+H]⁺, 100%).¹H NMR δ_(H) (400 MHz, D₆-DMSO): 3.85 (s, 3H), 5.93 (brs, 2H), 6.54 (t,1H), 7.23 (dd, 1H), 7.40 (d, 1H), 7.54 (d, 1H), 7.79 (d, 1H), 8.20 (d,1H).

Procedure for Synthesis of Compound 27.

A solution of sodium nitrite (3.14 g, 45.50 mmol, 1.0 eq.) in water (45mL) was slowly added to a stirred suspension of compound 26 (9.75 g,44.88 mmol, 1.0 eq.) in a mixture of concentrated aqueous HCl solution(45 mL) and water (45 mL) at 0° C. By the end of addition of the sodiumnitrite solution the reaction mixture became clear. After the additionformation of a precipitate was observed. The reaction mixture wasstirred at 0° C. for 10 minutes after the addition. Then a solution ofpotassium iodide (14.82 g, 89.28 mmol, 2.0 eq.) in water (45 mL) wasadded to the mixture at 0° C. A very viscous red-brown mixture wasformed which turned to dark brown color. The reaction mixture wasstirred at room temperature for 30 minutes, treated with saturatedaqueous potassium carbonate solution to reach pH>8 and extracted withDCM. The organic layer was washed with an aqueous NaHSO₃ solution, withwater, dried over sodium sulfate and concentrated at reduced pressure.The obtained residue was purified by column chromatography (silica gel,hexane/ethyl acetate, 4:1) giving compound 27 (12.04 g, 82%) as ayellowish solid. APCI-MS (m/z (intensity)): 329.07 ([M+H]⁺, 100%),370.09 ([M+MeCN+H]⁺, 20%).

Procedure for Synthesis of Compound 28.

Triethylamine (10 mL), t-Bu₃P (668 mg, 3.30 mmol, 9 mol %) andPdCl₂[PPh₃]₂ (773 mg, 1.10 mmol, 3 mol %) were added to a solution ofcompound 27 (12.02 g, 36.63 mmol, 1.0 eq.) in anhydrous DMF (60 mL). Theresulting mixture was stirred under argon atmosphere at room temperaturefor 10 minutes. Then phenyl-acetylene (7.49 g, 73.39 mmol, 2.0 eq.) wasadded dropwise. The reaction mixture was stirred at 75-80° C. for 2hours, cooled down to room temperature and filtered through a pad ofCelite washing with ethyl acetate. The filtrate was diluted with water(250 mL) and extracted with ethyl acetate. The organic phase was driedover sodium sulfate and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, hexane/DCM,4:1), then washed with cold diethyl ether and dried giving compound 28(8.68 g, 78%) as a yellowish solid. APCI-MS (m/z (intensity)): 303.18([M+H]⁺, 100%). ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 3.95 (s, 3H), 6.64 (t,1H), 7.43-7.47 (m, 3H), 7.51-7.55 (m, 2H), 7.86 (d, 1H), 7.89 (d, 1H),8.09 (dd, 1H), 8.24 (d, 1H), 8.62 (d, 1H).

Procedure for Synthesis of Compound 29.

A solution of NaOH (4.55 g, 113.75 mmol, 5.0 eq.) in water (40 mL) wasadded to a suspension of compound 28 (6.90 g, 22.82 mmol, 1.0 eq.) inMeOH (350 mL). The reaction mixture was stirred at 50-55° C. for 2.5hours, cooled down to room temperature, concentrated at reducedpressure, diluted with water (200 mL) and acidified with an aqueous HClsolution (1M) to reach pH 5. The formed precipitate was collected byfiltration, dried at reduced pressure with P₂O₅, washed with cold etherand dried giving compound 29 (6.38 g, 97%) as a yellowish solid. APCI-MS(m/z (intensity)): 289.12 ([M+H]⁺, 100%). ¹H NMR δ_(H) (400 MHz,D₆-DMSO): 3.50 (brs, 1H+H₂O), 6.58 (t, 1H), 7.40-7.44 (m, 3H), 7.46-7.50(m, 2H), 7.67 (d, 1H), 7.79 (d, 1H), 8.03 (dd, 1H), 8.19 (d, 1H), 8.48(d, 1H).

Procedure for Synthesis of Compound 30.

A mixture of compound 29 (4.97 g, 17.24 mmol, 1.0 eq.), TBTU (7.75 g,24.15 mmol, 1.4 eq.), DCM (50 mL) and THF (150 mL) was stirred at roomtemperature for 50 minutes. Then 3-imidazol-1-yl-propylamine (4) (2.38g, 18.98 mmol, 1.1 eq.) and DIPEA (6 mL, 34.50 mmol, 2.0 eq.) wereadded. The reaction mixture was stirred at room temperature for 2 hours,diluted with saturated aqueous NaHCO₃ solution (equivalent volume),stirred at room temperature for 1.5 hours and extracted with ethylacetate. The organic layer was dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, ethyl acetate/MeOH/NH₄OH, 40:2:1)giving compound 30 (5.78 g, 85%) as a white solid.

TABLE 1 Yield —NR1R2 Compound (%) 1

32a 60 2

32b 30 3

32c 36 4

32d 63 5

32e 60 6

32f 69 7

32g 31 8

32h 57 9

32i 70 10

32j 37 11

32k 74 12

32l 39 13

32m 65 14

32n 54 15

32o 74 16

32p 65 17

32q 37 18

32r 68 19

32s 75 20

32t 57 21

32u 80

Generic Procedure for Synthesis of Compounds 32a-u.

A suspension of compound 29 (140 mg, 0.48 mmol, 1.0 eq.), TBTU (263 mg,0.82 mmol, 1.7 eq.) and THF (5 mL) was stirred at room temperature for 2hours. Then the corresponding amine (R1R2NH) (0.58 mmol, 1.2 eq),triethylamine (0.2 mL, 1.44 mmol, 3.0 eq.) and DCM (3 mL) were added.The reaction mixture was stirred at room temperature for 4 hours,diluted with saturated aqueous NaHCO₃ solution (equivalent volume),stirred at room temperature for 2 hours and extracted with DCM. Theorganic layer was dried over sodium sulfate and concentrated at reducedpressure. The obtained residue was purified by column chromatographygiving a target compounds (32a-u).

Procedure for Synthesis of Compound 35.

A mixture of 3-bromo-benzoic acid (34) (700 mg, 3.48 mmol, 1.0 eq.),TBTU (1.680 g, 5.22 mmol, 1.5 eq.), 3-imidazol-1-yl-propylamine (4) (500mg, 3.99 mmol, 1.1 eq.), triethylamine (0.7 mL, 5.00 mmol, 1.4 eq.) andDCM (15 mL) was stirred at room temperature for 50 hours, diluted withsaturated aqueous NaHCO₃ solution (50 mL) and extracted with DCM. Theorganic layer was dried over sodium sulfate and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, ethyl acetate/MeOH), washed with diethyl ether and driedgiving compound 35 (855 mg, 80%) as a white solid.

Procedure for Synthesis of Compound 36.

A mixture of compound 35 (308 mg, 1.00 mmol, 1.0 eq.), PdCl₂[PPh₃]₂ (30mg, 0.04 mmol, 4 mol %), t-Bu₃P (30 mg, 0.15 mmol, 15 mol %),triethylamine (3 mL) and DMF (3 mL) was stirred under argon atmosphereat room temperature for 5 minutes. Then phenyl-acetylene (200 mg, 2.00mmol, 2.0 eq.) was added dropwise. The reaction mixture was stirred at75-80° C. for 2 hours, cooled down to room temperature, diluted withwater (20 mL) and extracted with DCM. The organic phase was dried oversodium sulfate and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, ethylacetate/MeOH/NH₄OH, 10:2:1) giving compound 36 (230 mg, 70%) as a whitesolid.

Experimental part: General experimental methods. LCMS. The LC/MSanalysis was done at Surveyor MSQ (Thermo Fisher Scientific) with APCIionization. 1. Type of HPLC column: Phenomenex Onyx Monolithic C18;25×4.6 mm; Part No: CHO-7645. 2. Solvent for samples dissolution: 50%DMSO, 50% acetonitrile. 3. Flow rate: 1.5 mL/min; column temperature 25°C. 4. Mobile phase: A=0.1% solution of formic acid in water, B=0.1%solution of formic acid in acetonitrile. 5. Gradient:

time, min. A % B % 0.0 100 0 0.1 100 0 2.1 5 95 2.5 5 95 2.6 100 0 4.0100 06. Detection: diode array (PDA), 200-800 nm; photodiode array detector.Detection was carried out in the full ultraviolet-visible range from 200to 800 nm. APCI (+ or/and − ions)—atmospheric pressure chemicalionization ELSD (PL-ELS 2100). 7. Total run time of the method: 4.5 min.8. Injection volume: 2 μL.

NMR: The ¹H NMR spectra were recorded on a MERCURY plus 400 MHzspectrometer (Varian). Chemical shift values are given in ppm relativeto tetramethylsilane (TMS), with the residual solvent proton resonanceas internal standard.

HPLC: The HPLC analysis was done at Agilent 1100 instrument. 1. Type ofHPLC column: Onyx Monolithic C18, 100×4.6 mm. 2. Flow rate: 1 mL/min;column temperature—ambient. 3. Mobile phase: A=0.1% TFA in water, B=0.1%TFA in acetonitrile.

List of abbreviations: Ac—acetyl, MeCO, APCI—atmospheric-pressurechemical ionization, aq.—aqueous, Ar—aryl or argon, atm—atmosphere(s),brs—broad singlet, Bu—butyl, conc.—concentrated, d—doublet,DABCO—1,4-diazabicyclo[2.2.2]octane, DCM—dichloromethane, dd—doublet ofdoublets, DIPEA—diisopropylethylamine, DMF—dimethylformamide,DMSO—dimethylsulfoxide, dppf—1,1′-bis(diphenylphosphino)ferrocene,ELSD—evaporative light scattering detector, Et—ethyl, eq.—equivalent,h—hour(s), HPLC—high-performance liquid chromatography, i-—iso-,i-Pr—i-propyl, m—multiplet, Me—methyl, MeCN—acetonitrile, MHz—megahertz,n-—normal-, n-Bu—n-butyl, min—minute(s), MS—mass-spectrometry,MWI—microwave irradiation, NBS—N-bromosuccinimide, NMR—Nuclear magneticresonance, PDA—photodiode array, Ph—phenyl, Pr—propyl, q—quartet,Ra—Ni—Raney-nickel, RT—room temperature, s—singlet, t—triplet, t-—tert-,TBTU—N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uroniumtetrafluoroborate, t-Bu—tert-butyl, THF—tetrahydrofuran, TMS(tms)—trimethylsilyl, UV—ultraviolet.

Example 2

This example provides synthetic procedures for benzamides andnicotinamides of the present disclosure.

TABLE 2 Product Yield Product —R compound (%) compound Yield (%) 1 3-Py—2a 96 3a 87 2 4-Me—C₆H₄— 2b 51 3b 88 3 4-F—C₆H₄— 2c 70 3c 94 44-CF₃—C₆H₄— 2d 81 3d 75 5 3-Cl—C₆H₄— 2e 76 3e 95 6 2-Cl—C₆H₄— 2f 84 3f96 7 2-CF₃—C₆H₄— 2g 84 3g 62 8 3-F—C₆H₄— 2h 56 3h 96 9 4-Cl—C₆H₄— 2i 793i 93 10 2-F—C₆H₄— 2j 73 3j 96 11 4-Py— 2k 97 3k 83 12 4-MeO—C₆H₄— 2l 423l 82 13 3-MeO—C₆H₄— 2m 75 3m 94 14 3-HO—C₆H₄— 2n 44 3n 97 15 2-Me—C₆H₄—2o 86 3o 89 16 3-Me—C₆H₄— 2p 59 3p 85 17 2-MeO—C₆H₄— 2q 49 3q 67 183-CF₃—C₆H₄— 2r 68 3r 82 19 —CMe₂OH 2s 91 3s 73 20

2t 97 3t 98 21

2u 38 3u 52 22 2-Py— 2v 64 3v 81 23

2w 62 3w 99 24

2x 52 3x 99 25

2y 73 3y 71 26

2z 35 3z 99 27 3-MeNHCO—C₆H₄— 2aa 61 3aa 99

TABLE 3 Starting Product Yield —R compound compound (%) 1 3-Py- 3a 5a 622 4-Me—C₆H₄— 3b 5b 65 3 4-F—C₆H₄— 3c 5c 69 4 4-CF₃—C₆H₄— 3d 5d 99 53-Cl—C₆H₄— 3e 5e 26 6 2-Cl—C₆H₄— 3f 5f 42 7 2-CF₃—C₆H₄— 3g 5g 88 83-F—C₆H₄— 3h 5h 53 9 4-Cl—C₆H₄— 3i 5i 69 10 2-F—C₆H₄— 3j 5j 66 11 4-Py-3k 5k 61 12 3-MeO—C₆H₄— 3m 5l 61 13 3-HO—C₆H₄— 3n 5m 42 14 3-Me—C₆H₄— 3p5n 81 15 3-CF₃—C₆H₄— 3r 5o 62

TABLE 4 Starting Product Yield —R compound compound (%) 1 3-Py— 3a 7a 592 4-Me—C₆H₄— 3b 7b 62 3 4-F—C₆H₄— 3c 7c 75 4 4-CF₃—C₆H₄— 3d 7d 70 53-Cl—C₆H₄— 3e 7e 62 6 2-Cl—C₆H₄— 3f 7f 58 7 2-CF₃—C₆H₄— 3g 7g 56 83-F—C₆H₄— 3h 7h 25 9 4-Cl—C₆H₄— 3i 7i 81 10 2-F—C₆H₄— 3j 7j 66 11 4-Py—3k 7k 46 12 4-MeO—C₆H₄— 3l 7l 54 13 3-MeO—C₆H₄— 3m 7m 68 14 3-HO—C₆H₄—3n 7n 47 15 2-Me—C₆H₄— 3o 7o 63 16 3-Me—C₆H₄— 3p 7p 62 17 2-MeO—C₆H₄— 3q7q 73 18 3-CF₃—C₆H₄— 3r 7r 54 19 —CMe₂OH 3s 7s 63 20

3t 7t 53 21

3u 7u 46 22 2-Py— 3v 7v 26 23

3w 7w 56 24

3x 7x 62 25

3y 7y 69 26

3z 7z 70 27 3-MeNHCO—C₆H₄— 3aa 7aa 49

Generic Procedure for Synthesis of Compounds 2a-aa:

To a solution of compound 1 (656 mg, 2.00 mmol, 1.0 eq.) in MeCN (10 mL)were added triethylamine (0.56 mL, 4.00 mmol, 2.0 eq.), then under argonatmosphere PdCl₂[PPh₃]₂ (70 mg, 0.10 mmol, 5 mol %), CuI (19 mg, 0.10mmol, 5 mol %) and the corresponding acetylene (3.00 mmol, 1.5 eq.). Themixture was refluxed under argon atmosphere for 4-8 hours, cooled downto room temperature and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, DCM/ethylacetate or hexane/ethyl acetate) to give a target compound (2a-aa).Compound 2a: yield 580 mg, 96% as a white solid. APCI-MS (m/z(intensity)): 304.20 ([M+H]′, 100%). Compound 2b: yield 320 mg, 51% as abeige solid. APCI-MS (m/z (intensity)): 317.22 ([M+H]⁺, 100%). Compound2c: yield 450 mg, 70% as a white solid. APCI-MS (m/z (intensity)):321.23 ([M+H]⁺, 100%). Compound 2d: yield 600 mg, 81% as a brown oil.Compound 2e: yield 514 mg, 76% as a white solid. Compound 2f: yield 565mg, 84% as a beige solid. Compound 2g: yield 625 mg, 84% as a beigesolid. Compound 2h: yield 270 mg, 56% as a white solid. APCI-MS (m/z(intensity)): 321.20 ([M+H]⁺, 100%). Compound 2i: yield 530 mg, 79% as awhite solid. APCI-MS (m/z (intensity)): 337.22, 338.44 ([M+H]⁺, 100%).Compound 2j: yield 465 mg, 73% as a white solid. Compound 2k: yield 586mg, 97% as a light-beige solid. APCI-MS (m/z (intensity)): 304.20([M+H]⁺, 100%). Compound 2l: yield 140 mg, 42% as a white solid. APCI-MS(m/z (intensity)): 330.10 ([M+H]⁺, 100%). Compound 2m: yield 500 mg, 75%as a white solid. APCI-MS (m/z (intensity)): 333.20 ([M+H]⁺, 100%).Compound 2n: yield 280 mg, 44% as a beige solid. Compound 2o: yield 540mg, 86% as a beige solid. Compound 2p: yield 370 mg, 59% as a colorlessoil. Compound 2q: yield 164 mg, 49% as a white solid. APCI-MS (m/z(intensity)): 333.06 ([M+H]⁺, 100%). Compound 2r: yield 500 mg, 68% as awhite solid. Compound 2s: yield 515 mg, 91% as a yellow oil. Compound2t: yield 630 mg, 97% as a yellow oil. Compound 2u: yield 240 mg, 38% asa yellow solid. APCI-MS (m/z (intensity)): 318.77 ([M+H]⁺, 100%).Compound 2v: yield 195 mg, 64% as a grey solid. Compound 2w: yield 200mg, 62% as a beige solid. Compound 2x: yield 175 mg, 52% as a beigesolid. Compound 2y: yield 250 mg, 73% as a brown solid. APCI-MS (m/z(intensity)): 343.12 ([M+H]⁺, 100%). Compound 2z: yield 120 mg, 35% as ayellow solid. APCI-MS (m/z (intensity)): 347.11 ([M+H]⁺, 100%). Compound2aa: yield 220 mg, 61% as a white solid.

Generic Procedure for Synthesis of Compounds 3a-aa:

The corresponding ester (2a-aa) (whole amount prepared on the previousstage) was dissolved in hot MeOH or MeOH-THF mixture (2:1, 10-25 mL).Then a solution of NaOH (200 mg, 5.00 mmol) in water (10 mL) was added.The resulting mixture was stirred at 50° C. for 1-2 hours (TLC control),cooled down to room temperature), acidified with concentrated aqueousHCl solution (to adjust pH 4-5). The formed precipitate was collected byfiltration, washed with cold water and diethyl ether and dried to give atarget compound (3a-aa). Compound 3a: yield 480 mg, 87% as a beigesolid. APCI-MS (m/z (intensity)): 290.11 ([M+H]⁺, 100%). APCI-MS (m/z(intensity)): 288.07 ([M−H]⁻, 85%), 334.12 ([M−H+formic acid]⁻, 100%).Compound 3b: yield 270 mg, 88% as a white solid. APCI-MS (m/z(intensity)): 303.17 ([M+H]⁺, 100%). APCI-MS (m/z (intensity)): 301.11([M−H]⁻, 100%), 347.19 ([M−H+formic acid]⁻, 90%). Compound 3c: yield 406mg, 94% as a white solid. APCI-MS (m/z (intensity)): 306.68 ([M+H]⁺,100%). APCI-MS (m/z (intensity)): 305.14 ([M−H]⁻, 100%), 651.22([M−H+formic acid]⁻, 85%). Compound 3d: yield 451 mg, 75% as abeige-green solid. APCI-MS (m/z (intensity)): 357.06 ([M+H]⁺, 100%).Compound 3e: yield 470 mg, 95% as a light-green solid. APCI-MS (m/z(intensity)): 323.09, 325.10 ([M+H]⁺, 100%). Compound 3f: yield 520 mg,96% as a white solid. APCI-MS (m/z (intensity)): 323.09, 325.09 ([M+H]⁺,100%). Compound 3g: yield 373 mg, 62% as a white solid. APCI-MS (m/z(intensity)): 357.07 ([M+H]⁺, 100%). Compound 3h: yield 248 mg, 96% as awhite solid. APCI-MS (m/z (intensity)): 305.13 ([M−H]⁻, 100%), 351.19([M−H+formic acid]⁻, 90%). Compound 3i: yield 470 mg, 93% as a whitesolid. APCI-MS (m/z (intensity)): 322.64, 323.84 ([M+H]⁺, 100%). APCI-MS(m/z (intensity)): 321.11, 322.35 ([M−H]⁻, 100%), 367.19 ([M−H+formicacid]⁻, 50%). Compound 3j: yield 426 mg, 96% as a white solid. APCI-MS(m/z (intensity)): 306.68 ([M+H]⁺, 100%). APCI-MS (m/z (intensity)):305.11 ([M−H]⁻, 100%), 351.20 ([M−H+formic acid]⁻, 60%). Compound 3k:yield 466 mg, 83% as a beige solid. APCI-MS (m/z (intensity)): 290.12([M+H]⁺, 100%). Compound 31: yield 110 mg, 82% as a grey solid. APCI-MS(m/z (intensity)): 319.21 ([M+H]⁺, 100%). APCI-MS (m/z (intensity)):317.13 ([M−H]⁻, 100%), 363.23 ([M−H+formic acid]⁻, 80%). Compound 3m:yield 448 mg, 94% as a white solid. APCI-MS (m/z (intensity)): 319.23([M+H]⁺, 100%). Compound 3n: yield 260 mg, 97% as a beige solid. APCI-MS(m/z (intensity)): 305.18 ([M+H]⁺, 100%). APCI-MS (m/z (intensity)):303.13 ([M−H]⁻, 45%), 349.17 ([M−H+formic acid]⁻, 100%). Compound 3o:yield 460 mg, 89% as a white solid. APCI-MS (m/z (intensity)): 302.51([M+H]⁺, 100%). Compound 3p: yield 300 mg, 85% as a white solid. APCI-MS(m/z (intensity)): 302.45 ([M+H]⁺, 100%). Compound 3q: yield 105 mg, 67%as a white solid. APCI-MS (m/z (intensity)): 319.07 ([M+H]⁺, 100%).Compound 3r: yield 386 mg, 80% as a light-green solid. APCI-MS (m/z(intensity)): 357.07 ([M+H]⁺, 100%). Compound 3s: yield 357 mg, 73% as abeige solid. APCI-MS (m/z (intensity)): 270.45 ([M+H]⁺, 100%), 253.10([M−H₂O+H]⁺, 65%). Compound 3t: yield 590 mg, 98% as a white solid.APCI-MS (m/z (intensity)): 311.19 ([M+H]⁺, 70%), 293.17 ([M−H₂O+H]⁺,100%). Compound 3u: yield 120 mg, 52% as a beige solid. APCI-MS (m/z(intensity)): 305.13 ([M+H]⁺, 100%). Compound 3v: yield 150 mg, 81% as agrey solid. APCI-MS (m/z (intensity)): 290.14 ([M+H]⁺, 100%). Compound3w: yield 190 mg, 99% as a beige solid. APCI-MS (m/z (intensity)):308.13 ([M+H]⁺, 100%). Compound 3x: yield 165 mg, 99% as a beige solid.APCI-MS (m/z (intensity)): 324.11, 326.10 ([M+H]⁺, 100%). APCI-MS (m/z(intensity)): 322.01 ([M−H]⁺, 100%), 367.96 ([M−H+formic acid]⁻, 60%).Compound 3y: yield 170 mg, 71% as a beige solid. APCI-MS (m/z(intensity)): 329.35 ([M+H]⁺, 100%). Compound 3z: yield 115 mg, 99% as abeige solid. APCI-MS (m/z (intensity)): 33.07 ([M+H]⁺, 100%). Compound3aa: yield 210 mg, 99% as a white solid. APCI-MS (m/z (intensity)):346.10 ([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 5a-o:

A mixture of the corresponding acid derivative (3a-k, m,n,p,r)(0.33-0.86 mmol, 1.0 eq.), TBTU (1.2 eq.), triethylamine (3.0 eq.) anddry DMF (5 mL) was stirred at room temperature for 5 minutes. Then3-imidazol-1-yl-propylamine (4) (1.2 eq.) was added. The resultedmixture was stirred at room temperature for 8-12 hours, diluted withwater (100 mL), extracted with ethyl acetate (3×50 mL). The organiclayers were combined, washed with an aqueous K₂CO₃ solution (30 mL),water (3×30 mL), dried over sodium sulfate and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, DCM/MeOH) to give a target compound (5a-o). Compound 5a:yield 197 mg, 62% as a white solid. Compound 5b: yield 115 mg, 65% as awhite solid. Compound 5c: yield 186 mg, 69% as a white solid. Compound5d: yield 290 mg, 99% as a white solid. Compound 5e: yield 80 mg, 26% asa white solid. Compound 5f: yield 145 mg, 42% as a white solid. Compound5g: yield 213 mg, 88% as a white solid. Compound 5h: yield 86 mg, 53% asa grey solid. Compound 5i: yield 210 mg, 69% as a white solid. Compound5j: yield 187 mg, 66% as a white solid. Compound 5k: yield 193 mg, 61%as a beige solid. Compound 5l: yield 180 mg, 61% as a white solid.Compound 5m: yield 70 mg, 42% as a white solid. Compound 5n: yield 167mg, 81% as a white solid. Compound 5o: yield 153 mg, 62% as a whitesolid.

Generic Procedure for Synthesis of Compounds 7a-aa:

The same as Generic procedure for synthesis of compounds 5a-o using3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (1.2 eq.) andtriethylamine (4.0 eq.). Compound 7a: yield 188 mg, 59% as a whitesolid. Compound 7b: yield 109 mg, 62% as a beige solid. Compound 7c:yield 200 mg, 75% as a white solid. Compound 7d: yield 204 mg, 70% as awhite solid. Compound 7e: yield 188 mg, 62% as a white solid. Compound7f: yield 200 mg, 58% as a white solid. Compound 7g: yield 134 mg, 56%as a white solid. Compound 7h: yield 40 mg, 25% as a colorless oil.Compound 7i: yield 246 mg, 81% as a white solid. Compound 7j: yield 188mg, 66% as a yellowish oil. Compound 7k: yield 145 mg, 46% as a beigesolid. Compound 7l: yield 80 mg, 54% as a beige solid. Compound 7m:yield 200 mg, 68% as a white solid. Compound 7n: yield 78 mg, 47% as awhite solid. Compound 7o: yield 85 mg, 63% as a white solid. Compound7p: yield 128 mg, 62% as a white solid. Compound 7q: yield 102 mg, 73%as a white solid. Compound 7r: yield 133 mg, 54% as a white solid.Compound 7s: yield 87 mg, 63% as a beige solid. Compound 7t: yield 75mg, 53% as a white solid. Compound 7u: yield 75 mg, 46% as a whitesolid. Compound 7v: yield 53 mg, 26% as a brown oil. Compound 7w: yield143 mg, 56% as a beige solid. Compound 7x: yield 136 mg, 62% as a beigesolid. Compound 7y: yield 155 mg, 69% as a white solid. Compound 7z:yield 108 mg, 70% as a white solid. Compound 7aa: yield 135 mg, 49% as awhite solid.

Procedure for Synthesis of Compound 9.

To a solution of compound 1 (656 mg, 2.00 mmol, 1.0 eq.) in MeOH (20 mL)was added a solution of NaOH (200 mg, 5.00 mmol, 2.5 eq.) in water (10mL) and the reaction mixture was stirred at 50° C. for 1 hour, cooleddown to room temperature, acidified with concentrated aqueous HClsolution to adjust pH 4-5. The formed precipitate was collected byfiltration, washed with cold water and diethyl ether and dried to givecompound 9 (580 mg, 92%) as a white solid. APCI-MS (m/z (intensity)):315.00 ([M+H]⁺, 100%). APCI-MS (m/z (intensity)): 312.94 ([M−H]⁻, 100%),358.91 ([M−H+formic acid]⁻, 80%).

Procedure for Synthesis of Compound 10.

A mixture of compound 9 (580 mg, 1.85 mmol, 1.0 eq.), TBTU (709 mg, 2.20mmol, 1.2 eq.), triethylamine (0.98 mL, 7.00 mmol, 3.4 eq.) and dry DMF(30 mL) was stirred at room temperature for 5 minutes. Then3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (356 mg, 2.20 mmol,1.2 eq.) was added. The resulted mixture was stirred at room temperaturefor 8 hours, diluted with water (300 mL) and extracted with ethylacetate (3×150 mL). The organic layers were combined, washed with anaqueous K₂CO₃ solution (100 mL), water (3×100 mL), dried over sodiumsulfate and concentrated at reduced pressure. The obtained residue waspurified by column chromatography (silica gel, DCM/MeOH) to givecompound 10 (770 mg, 99%) as a white solid. APCI-MS (m/z (intensity)):422.02 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 13.

To a solution of compound 10 (210 mg, 0.50 mmol, 1.0 eq.) in a mixtureof DCM (10 mL) and THF (5 mL) were added triethylamine (0.1 mL, 0.71mmol, 1.4 eq.) and Boc₂O (125 mg, 0.58 mmol, 1.16 eq.). The mixture wasstirred at room temperature for 4 hours and concentrated at reducedpressure to give crude product 11 as colorless oil, which was used onthe next step without purification. The crude product 11 (approx. 0.50mmol, 1.0 eq.) was dissolved in MeCN (20 mL). Then triethylamine (0.4mL), PdCl₂[PPh₃]₂ (10 mg, 0.015 mmol, 3 mol %), CuI (10 mg, 0.05 mmol,10 mol %) and 2-ethynyl-6-pyrrolidin-1-yl-pyridine (12) (125 mg, 0.73mmol, 1.46 eq.) were added under argon atmosphere. The reaction mixturewas refluxed for 4 hours, cooled down to room temperature andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, DCM/ethyl acetate, 2:1) to givecompound 13 (112 mg, 40%) as a colorless oil. APCI-MS (m/z (intensity)):566.53 ([M+H]⁺, 100%), 466.43 ([M−Boc+H]⁺, 20%).

Procedure for Synthesis of Compound 14.

To a solution of compound 13 (112 mg, 0.20 mmol, 1.0 eq.) in MeOH (10mL) was added an aqueous HCl solution (15%, 2 mL). The resulted mixturewas stirred at room temperature for 1 hour, neutralized with an aqueousK₂CO₃ solution and extracted with DCM (2×50 mL). The combined organiclayers were dried over sodium sulfate and concentrated. The obtainedresidue was purified by column chromatography (silica gel, DCM/MeOH,20:1) to give compound 14 (43 mg, 47%) as a beige solid.

Procedure for Synthesis of Compound 16.

A mixture of compound 1 (3.28 g, 10.00 mmol, 1.0 eq.), triethylamine(4.2 mL, 30.00 mmol, 3.0 eq.), PdCl₂[PPh₃]₂ (0.35 g, 0.50 mmol, 5 mol%), CuI (0.19 g, 1.00 mmol, 10 mol %) and TMS-acetylene (1.96 g, 20.00mmol, 2.0 eq.) and MeCN (25 mL) was refluxed for 3.5 hours under argonatmosphere and concentrated at reduced pressure. The obtained residuewas purified by column chromatography (silica gel, hexane/ethyl acetate,10:1) to give compound 16 (1.90 g, 64%) as a yellow oil. APCI-MS (m/z(intensity)): 299.15 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 17.

To a solution of compound 16 (1.90 g, 6.38 mmol, 1.0 eq.) in hexane (50mL) was added dropwise a solution of TBAF trihydrate (0.85 g, 3.19 mmol,0.50 eq.) in ethyl acetate (10 mL) at 0° C. The reaction mixture wasstirred for 20 minutes at 0° C., washed with water (2×20 mL), dried oversodium sulfate and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, DCM) to givecompound 17 (1.11 g, 77%) as a yellowish solid. APCI-MS (m/z(intensity)): 227.16 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 18.

To a solution of compound 17 (470 mg, 2.08 mmol, 1.0 eq.) in MeOH (10mL) was added a solution of NaOH (200 mg, 5.00 mmol, 2.4 eq.) in water(10 mL). The resulted mixture was stirred at 40° C. for 1 hour, cooleddown to room temperature, acidified with concentrated aqueous HClsolution to adjust pH 4-5. The formed precipitate was collected byfiltration, washed with cold water and diethyl ether and dried to givecompound 18 (435 mg, 99%) as a beige solid. APCI-MS (m/z (intensity)):213.19 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 19.

Compound 19 was prepared according to Generic procedure for synthesis ofcompounds 5a-o using compound 18 (210 mg, 0.99 mmol). Yield 78 mg, 25%as a beige solid.

Procedure for Synthesis of Compound 20.

Compound 20 was prepared according to Generic procedure for synthesis ofcompounds 7a-aa using compound 18 (210 mg, 0.99 mmol). Yield 144 mg, 46%as a white solid.

Procedure for Synthesis of Compound 22.

Compound 22 was prepared according to Generic procedure for synthesis ofcompounds 2a-aa using compound 1 (328 mg, 1.00 mmol, 1.0 eq.) and(4-ethynyl-benzyl)-carbamic acid tert-butyl ester (21) (300 mg, 1.30mmol, 1.3 eq.). Yield 188 mg, 44% as a yellowish solid.

Procedure for Synthesis of Compound 23.

Compound 23 was prepared according to Generic procedure for synthesis ofcompounds 3a-aa using compound 22 (188 mg, 0.44 mmol). Yield 177 mg, 97%as a beige solid. APCI-MS (m/z (intensity)): 418.19 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 24.

Compound 24 was prepared according to Generic procedure for synthesis ofcompounds 7a-aa using compound 23 (177 mg, 0.42 mmol). Yield 133 mg, 50%as a white solid.

Procedure for Synthesis of Compound 25.

To a solution of compound 24 (93 mg, 0.18 mmol, 1.0 eq.) in MeOH (5 mL)was added an aqueous HCl solution (15%, 2 mL). The reaction mixture wasstirred at room temperature for 8 hours, neutralized with an aqueousK₂CO₃ solution and extracted with DCM (2×50 mL). The combined organiclayers were dried over sodium sulfate and concentrated. The obtainedresidue was purified by column chromatography (silica gel, DCM/MeOH,10:1) to give compound 25 (70 mg, 92%) as a beige solid

Procedure for Synthesis of Compound 27.

Compound 27 was prepared according to Generic procedure for synthesis ofcompounds 2a-aa using compound 1 (656 mg, 2.00 mmol, 1.0 eq.) and4-ethynyl-benzonitrile (26) (380 mg, 3.00 mmol, 1.5 eq.). Yield 437 mg,67% as a beige-yellow solid. APCI-MS (m/z (intensity)): 328.14 ([M+H]⁺,100%), 369.11 ([M+MeCN+H]⁺, 100%).

Procedure for Synthesis of Compounds 30 and 31.

A mixture of intermediates 28 and 29 (420 mg, with 2:3: ratio accordingto LCMS) was prepared according to Generic procedure for synthesis ofcompounds 3a-aa using compound 27 (437 mg, 1.34 mmol). The obtainedmixture was used for next step without separation. A part of the mixture(150 mg, approx. 0.48 mmol), TBTU (225 mg, 0.70 mmol, 1.5 eq.),triethylamine (0.28 mL, 2.0 mmol, 4.2 eq.) and DMF dry (5 mL) werestirred at room temperature for 5 minutes. Then3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (113 mg, 0.70 mmol,1.5 eq.) was added. The resulted mixture was stirred at room temperaturefor 12 hours, diluted with water (100 mL) and extracted with ethylacetate (3×50 mL). The organic layers were combined, washed with anaqueous K₂CO₃ solution (30 mL), water (3×30 mL), dried over sodiumsulfate and concentrated at reduced pressure. The obtained residue waspurified by column chromatography (silica gel, DCM/MeOH) to giveseparated compounds 30 (76 mg, 38% for 2 steps) as a white solid and 31(63 mg, 30% for 2 steps) as a white solid.

Procedure for Synthesis of Compound 34.

A mixture of compound 33 (300 mg, 0.76 mmol, 1.0 eq.), 10% Pd/C catalyst(100 mg, 0.094 mmol, 9 mol %) and MeOH (50 mL).) was stirred underhydrogen atmosphere at room temperature for 3 hours. The catalyst wasremoved by filtration. The filtrate was concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, ethyl acetate/MeOH/NH₄OH, 40:2:1) giving compound 34 (252mg, 83%) as a white solid.

Procedure for Synthesis of Compound 36.

A mixture of 4-(1H-pyrazol-1-yl)benzoic acid (35) (278 mg, 1.48 mmol,1.00 eq.), 3-(1H-imidazol-1-yl)propan-1-amine (4) (200 mg, 1.60 mmol,1.08 eq.), TBTU (622 mg, 2.00 mmol, 1.35 eq.), triethylamine (0.28 mL,2.00 mmol, 1.35 eq.) and DCM (10 mL) was stirred at room temperature for20 hours, diluted with equal volume of saturated aqueous NaHCO₃ solutionand stirred at room temperature for 2 hours. The resulting mixture wasextracted with DCM. The organic phase was dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, ethyl acetate/MeOH, 20:1), then wasrecrystallized from DCM/hexane giving compound 36 (101 mg, 23%) as ayellow solid.

TABLE 5 Condi- Yield Condi- Yield Yield —NR1R2 Comp. tions (%)^(a) Comp.tions (%) Comp. (%) 1

43a a, c 91 44a d 78 45a 99 2

43b a, c 75 44b d 70 45b 92 3

43c a, c 95 44c d 70 45c 93 4

43d a, c 84 44d d 59 45d 93 5

43e a, c 60 44e d 71 45e 94 6

43f b, c 48 44f d 56 45f 91 7

43g a, c 86 44g e 30 45g 49 ^(a)yield for 2 steps

TABLE 6 Starting Product Condi- Yield —NR1R2 compound compound tions (%)1

45a 46a g 99 2

45b 46b g 79 3

45c 46c g 70 4

45d 46d g 80 5

45e 46e g 89 6

45f 46f g 78 7

46g 46g j 64

TABLE 7 Starting Product Yield —NR1R2 compound compound (%) 1

46a 49a 38 2

46b 49b 78 3

46c 49c 98 4

46d 49d 56 5

46e 49e 90 6

46f 49f 61 7

46g 49g 39 8

46h 49h 48

Generic Procedure for Synthesis of Compounds 43a-e.

A mixture of 4-chloro-3-nitro-benzoic acid methyl ester (41) (5.39 g,25.00 mmol, 1.0 eq.), corresponding amine (37.50 mmol, 1.5 eq.), DIPEA(4.35 mL, 25.00 mmol, 1.0 eq.) and ethanol (50 mL) was heated at refluxfor 6 hours, then cooled to room temperature, and concentrated atreduced pressure. The obtained residue was purified by columnchromatography (silica gel, ethyl acetate/hexane) to give anintermediate (42a-e) used on the next stage without additionalcharacterization. Nickel powder (10% weight to an intermediate 42a-e)was added to a solution of the corresponding intermediate (42a-e) inmethanol. The obtained mixture was stirred at room temperature under H₂(3-4 atm) for 8 hours, filtered and concentrated at reduced pressure.The obtained residue was purified by column chromatography (silica gel,ethyl acetate/hexane) to give a target compound (43a-e). Compound 43a:yield 5.33 g, 91% (for 2 steps) as a yellow solid. APCI-MS (m/z(intensity)): 235.23 ([M+H]⁺, 100%). Compound 43b: yield 4.63 g, 75%(for 2 steps) as a yellow solid. APCI-MS (m/z (intensity)): 279.20([M+H]⁺, 100%). Compound 43c: yield 2.15 g, 95% (for 2 steps) as ayellow solid. APCI-MS (m/z (intensity)): 456.08 ([M+H]⁺, 100%). Compound43d: yield 6.04 g, 84% (for 2 steps) as a yellowish solid. APCI-MS (m/z(intensity)): 289.21 ([M+H]⁺, 100%). Compound 43e: yield 0.51 g, 30%(for 2 steps) as a yellowish solid. APCI-MS (m/z (intensity)): 302.80([M+H]⁺, 100%).

Procedure for Synthesis of Compound 43f.

A mixture of 4-chloro-3-nitro-benzoic acid methyl ester (41) (5.39 g,25.00 mmol, 1.0 eq.), 3-hydroxymethylpiperidine (3.23 g, 25.00 mmol, 1.0eq.), DIPEA (3.51 mL, 25.00 mmol, 1.0 eq.) and DMF (50 mL) was heated at100° C. for 5 hours, then the second portion of3-hydroxymethylpiperidine (323 mg, 2.50 mmol, 0.1 eq.) was added. Themixture was stirred at 100° C. for 8 hours, cooled to room temperature,poured to water and extracted with ethyl acetate. The organic phase waswashed with brine and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, ethylacetate/hexane) to give the intermediate 42f as a yellow oil (4.13 g)used on the next stage without additional characterization. Nickelpowder (413 mg, 10% weight) was added to a solution of the intermediate42f in methanol (10%, 40 mL). The obtained mixture was stirred at roomtemperature under H₂ (3-4 atm) for 8 hours filtered and concentrated atreduced pressure. The obtained residue was purified by columnchromatography (silica gel, ethyl acetate/hexane) to give compound 43fas a yellow oil (3.34 g, 48% for 2 steps). APCI-MS (m/z (intensity)):279.21 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 43g.

A mixture of 4-chloro-3-nitro-benzoic acid methyl ester (41) (5.39 g,25.00 mmol, 1.0 eq.), N-boc-piperazine (5.12 g, 27.50 mmol, 1.1 eq.),DIPEA (4.35 mL, 25.00 mmol, 1.0 eq.) and ethanol (50 mL) was heated atreflux for 4 hours, then cooled to room temperature, and concentrated atreduced pressure. The obtained residue was purified by chromatography(silica gel, ethyl acetate/hexane) to give the intermediate 42g as ayellowish oil (8.77 g). Nickel powder (877 mg, 10% weight) was added toa solution of the intermediate 42g in methanol (10%, 90 mL). Theobtained mixture was stirred at room temperature under H₂ (3-4 atm) for8 hours, filtered and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, ethylacetate/hexane) to give compound 43g as a yellow solid (7.21 g, 86% for2 steps). APCI-MS (m/z (intensity)): 336.18 ([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 44a-f:

A solution of sodium nitrite (1.58 g, 23.00 mmol, 1.02 eq.) in water (23mL) was slowly added to a stirred solution of the corresponding amine(43a-f) (22.62 mmol, 1.00 eq.) in a mixture of concentrated aqueous HClsolution (23 mL) and water (23 mL) at −5° C. By the end of addition ofthe sodium nitrite solution the reaction mixture became clear. Thereaction mixture was stirred at −5° C.÷−2° C. for 10 minutes after theaddition. Then a solution of potassium iodide (7.51 g, 43.00 mmol, 1.90eq.) in water (23 mL) was added to the mixture at −2° C. Then thereaction mixture was stirred at room temperature for 30 minutes, dilutedwith DCM, treated with saturated aqueous potassium carbonate solution toreach pH>8 and extracted with DCM. The organic layer was washed with anaqueous Na₂S₂O₅ solution, with water, dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, ethyl acetate/hexane) to give atarget compound (44a-f). Compound 44a: yield 6.11 g, 78% as a red oil.APCI-MS (m/z (intensity)): 346.04 ([M+H]⁺, 100%). Compound 44b: yield4.70 g, 70% as a yellow oil. APCI-MS (m/z (intensity)): 360.02 ([M+H]⁺,100%). Compound 44c: yield 2.16 g, 70% as reddish oil. APCI-MS (m/z(intensity)): 374.04 ([M+H]⁺, 100%). Compound 44d: yield 4.97 g, 59% asred solid. APCI-MS (m/z (intensity)): 400.00 ([M+H]⁺, 100%). Compound44e: yield 0.50 g, 71% as an orange oil. APCI-MS (m/z (intensity)):413.59 ([M+H]⁺, 100%). Compound 44f: yield 2.60 g, 56% as a red oil.APCI-MS (m/z (intensity)): 390.04 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 44g.

A solution of sodium nitrite (1.51 g, 21.93 mmol, 1.05 eq.) in water (23mL) was slowly added to a stirred solution of compound 43g (7.21 g,21.50 mmol, 1.00 eq.) in a mixture of concentrated aqueous HCl solution(23 mL) and water (23 mL) at −8° C. By the end of the addition of thesodium nitrite solution the reaction mixture became clear. The reactionmixture was stirred at −8° C. to −2° C. for 10 minutes after theaddition. Then a solution of potassium iodide (7.14 g, 43.00 mmol, 2.00eq.) in water (23 mL) was added to the mixture at −2° C. Then thereaction mixture was stirred at room temperature for 30 minutes, dilutedwith DCM, treated with saturated aqueous potassium carbonate solution toreach pH>8 and extracted with DCM. The organic layer was washed with anaqueous Na₂S₂O₅ solution, water, dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, ethyl acetate/hexane) to yield amixture of compound 44g and end a byproduct—a derivative of compound 44gwith cleaved Boc protecting group (1:1, 2.03 g). The resulted mixturewas dissolved in EtOH (30 mL). Boc₂O (0.5 eq.) was added, and themixture was stirred at room temperature for 16 hours, poured to waterand extracted with ethyl acetate. The organic phase was concentrated.The obtained residue was purified by column chromatography (silica gel,ethyl acetate/hexane) to give compound 44g as a colorless oil (2.85 g,30%). APCI-MS (m/z (intensity)): 446.59 ([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 45a-g.

A mixture of the corresponding iodide (44a-g) (3.24 mmol, 1.0 eq.),4-fluorophenylacetylene (0.584 g, 4.86 mmol, 1.5 eq.),PdCl₂[PPh₃]₂(0.070 g, 0.10 mmol, 3 mol %), copper iodide (0.019 g, 0.10mmol, 3 mol %), triethylamine (0.910 mL, 6.48 mmol, 2.0 eq.) andanhydrous acetonitrile (20 mL) was stirred under argon atmosphere at 60°C. for 1.5 hours, cooled to room temperature, poured to water andextracted with ethyl acetate. The organic phase was, washed with brineand concentrated at reduced pressure. The obtained residue was purifiedby column chromatography (silica gel, ethyl acetate/hexane) to give atarget compound (45a-g). Compound 45a: yield 1.08 g, 99% as a yellowishoil. APCI-MS (m/z (intensity)): 338.16 ([M+H]⁺, 100%). Compound 45b:yield 0.58 g, 92% as a yellowish oil. APCI-MS (m/z (intensity)): 352.16([M+H]⁺, 100%). Compound 45c: yield 0.51 g, 93% as a yellowish oil.APCI-MS (m/z (intensity)): 365.68 ([M+H]⁺, 100%). Compound 45d: yield0.55 g, 93% as a yellowish oil. APCI-MS (m/z (intensity)): 392.22([M+H]⁺, 100%). Compound 45e: yield 0.43 g, 94% as a yellowish oil.APCI-MS (m/z (intensity)): 406.14 ([M+H]⁺, 100%). Compound 45f: yield0.52 g, 91% as a yellowish oil. APCI-MS (m/z (intensity)): 439.19([M+H]⁺, 100%). Compound 45g: yield 1.36 g, 49% as a yellowish oil.APCI-MS (m/z (intensity)): 439.19 ([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 46a-f

To a solution of the corresponding ester (45a-f) (1.66 mmol, 1.0 eq.) inTHF (4 mL) and MeOH (4 mL) an aqueous solution of sodium hydroxide (50%,250 μL, 5.00 mmol, 3.0 eq.) was added, and the reaction mixture wasstirred at room temperature for 16 hours, concentrated at reducedpressure, diluted with water, treated with an aqueous HCl solution (1M)to reach pH 5 and extracted with ethyl acetate. The combined organiclayers were washed with water, and concentrated at reduced pressure. Theobtained residue was purified by chromatography (silica gel,ethanol/DCM) to give a target compound (46a-f). Compound 46a: yield 1.08g, 99% as a beige solid. APCI-MS (m/z (intensity)): 324.18 ([M+H]⁺,100%). Compound 46b: yield 0.44 g, 79% as a brown solid. APCI-MS (m/z(intensity)): 338.19 ([M+H]⁺, 100%). Compound 46c: yield 0.35 g, 70% asa brownish solid. APCI-MS (m/z (intensity)): 352.18 ([M+H]⁺, 100%).Compound 46d: yield 0.42 g, 80% as a brownish solid. APCI-MS (m/z(intensity)): 378.24 ([M+H]⁺, 100%). Compound 46e: yield 0.37 g, 89% asa brownish solid. APCI-MS (m/z (intensity)): 392.13 ([M+H]⁺, 100%).Compound 46f: yield 0.39 g, 78% as a white solid. APCI-MS (m/z(intensity)): 368.12 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 46g.

To a solution of compound 45g (1.088 g, 2.48 mmol, 1.0 eq.) in THF (15mL) and MeOH (18 mL) a solution of LiOH*H₂O (420 mg, 10.00 mmol, 4.0eq.) in water (16 mL) was added, and the reaction mixture was stirred atroom temperature for 16 hours, diluted with water, treated with anaqueous solution of HCl to reach pH 5 and extracted with ethyl acetate.The combined organic layers were washed with water, and concentrated atreduced pressure. The obtained residue was purified by columnchromatography (silica gel, ethanol/DCM) to give compound 46g (676 mg,64%) as a beige solid. APCI-MS (m/z (intensity)): 425.16 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 48.

A solution of compound 45g (272 mg, 0.62 mmol, 1.0 eq.) in THF (5 mL)was treated with a solution of HCl in dioxane (16%, 4.7 mL). Thereaction mixture was stirred at room temperature for 21 hour, poured toan aqueous NaHCO₃ solution and extracted with ethyl acetate. The organicphase was concentrated at reduced pressure. The obtained residue wasdried giving the intermediate 47 (238 mg) as a beige solid, which wasused on the next stage without additional purification andcharacterization. The intermediate 47 (238 mg), HCO₂H (380 μL, 10.00mmol), paraform (93 mg, 3.10 mmol) and EtOH (3 mL) was stirred at refluxfor 5 hours, then an aqueous solution of formaldehyde (40%, 430 μL) wasadded. The resulted mixture was stirred at reflux for 9 hours, cooled toroom temperature, poured to an aqueous NaHCO₃ solution and extractedwith ethyl acetate. The organic phase was concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, ethanol/DCM) to give of compound 48 (81 mg, 37% for 2steps) as dark brown oil. APCI-MS (m/z (intensity)): 353.16 ([M+H]⁺,100%).

Procedure for Synthesis of Compound 46h.

To a solution of compound 48 (81 mg, 0.23 mmol, 1.0 eq.) in MeOH (4 mL)a solution of LiOH*H₂O (28 mg, 0.69 mmol, 3.0 eq.) in water (4 mL) wasadded, and the reaction mixture was stirred at room temperature for 16hours, diluted with water, treated with an aqueous HCl solution to reachpH 6 and extracted with ethyl acetate. The combined organic layers werewashed with water and concentrated at reduced pressure. The obtainedresidue was dried giving compound 46h (58 mg, 75%) as a white solid.APCI-MS (m/z (intensity)): 339.18 ([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 49a-h.

A mixture of the corresponding acid (46a-h) (0.40 mmol, 1.0 eq.), TBTU(180 mg, 0.56 mmol, 1.4 eq.), triethylamine (0.126 μL, 0.90 mmol, 2.2eq.) and DMF (3 mL) was stirred at room temperature for 30 minutes. Then3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (78 mg, 0.48 mmol, 1.2eq.) was added. The reaction mixture was stirred at room temperature for5-16 hours, poured to an aqueous NaOH solution (1N) and extracted withethyl acetate. The organic phase was washed with water and concentrated.The obtained residue was purified by column chromatography (silica gel,ethanol/DCM) to give a target compound (49a-h). Compound 49a: yield 41mg, 38% as a yellow solid. Compound 49b: yield 139 mg, 78% as a yellowsolid. Compound 49c: yield 181 mg, 98% as a white solid. Compound 49d:yield 110 mg, 56% as a yellow solid. Compound 49e: yield 179 mg, 90% asa yellow solid. Compound 49f: yield 146 mg, 61% as a white solid.Compound 49g: yield 67 mg, 39% as a white solid Compound 49h: yield 36mg, 48% as a white solid

Procedure for Synthesis of Compound 50.

A solution of compound 49g (52 mg, 0.098 mmol) in THF (5 mL) was treatedwith a solution of HCl in dioxane (16%, 0.5 mL) and stirred at roomtemperature for 16 hours, poured to an aqueous NaHCO₃ solution, dilutedwith EtOH, and extracted with ethyl acetate. The organic phase wasconcentrated at reduced pressure. The obtained residue was dried givingcompound 50 (48 mg, 99%) as a beige solid.

TABLE 8 Starting Product Condi- Yield comp. R1 R2 compound R3 tions (%)1 41 Cl Me 53a Me₂N— a 93 2 52a Cl Et 53b

a 66 3 52a Cl Et 53c

a 99 4 41 Cl Me 53d PhO— b 70 5 52b F Et 53e

c 90 6 52b F Et 53f

c 86 7 41 Cl Me 53g^(a) 3-Py— d 43 8 41 Cl Me 53h^(a) 4-Py d 34 9 41 ClMe 53i^(a)

d 70 ^(a)compounds were obtained as a Et esters

TABLE 9 Starting Product Yield Product Yield R3 R2 comp. comp. (%) comp.(%) 1 Me₂N— Me 53a 54a 59 55a 25 2

Et 53b 54b 51 55b 70 3

Et 53c 54c 46 55c 65 4 PhO— Me 53d 54d 93 55d 50 5

Et 53e 54e 93 55e 22 6

Et 53f 54f 72 55f 77 7 3-Py— Et 53g 54g 60 55g 56 8 4-Py Et 53h 54h 9255h 59 9

Et 53i 54i 90 55i 81

TABLE 10 Starting Condi- Product Yield Product Yield R3 R2 comp. R4tions comp. (%) comp. (%) 1 Me₂N— Me 55a H a 56a 83 57a 48 2

Et 55b H a 56b 72 57b 60 3

Et 55c H a 56c 89 57c 90 4 PhO— Me 55d H a 56d 78 57d 15 5

Et 55e H a 56e 67 57e 83 6

Et 55e F b 56f 72 57f 95 7

Et 55f H a 56g 85 57g 94 8

Et 55f F b 56h 95 57h 89 9 3-Py— Et 55g H a 56i 92 57i 94 10 4-Py Et 55hH a 56j 95 57j 96 11 4-Py Et 55h F b 56k 80 57k 90 12

Et 55i F b 56l 70 57i 95

TABLE 11 Starting Product Yield R3 R4 comp. comp. (%) 1 Me₂N— H 57a 58a69 2

H 57b 58b 80 3

H 57c 58c 79 4 PhO— H 57d 58d 80 5

H 57e 58e 90 6

H 57g 58f 43 7 4-Py H 57j 58g 60

TABLE 12 Starting Product Yield R3 R4 comp. comp. (%) 1 Me₂N— H 57a 59a41 2

H 57b 59b 18 3

H 57c 59c 52 4 PhO— H 57d 59d 42 5

F 57f 59e 35 6

H 57g 59f 37 7

F 57h 59g 52 8 3-Py— H 57i 59h 53 9 4-Py H 57j 59i 28 10 4-Py F 57k 59j48 11

F 57l 59k 43

Generic Procedure for Synthesis of Compounds 53a-c.

A suspension of 4-chloro-3-nitro-benzoic acid methyl ester (41) or4-chloro-3-nitro-benzoic acid ethyl ester (52a) (31.57 mmol, 1.0 eq.),the corresponding amine (35.00 mmol, 1.1 eq.) and triethylamine (5.6 mL,40.00 mmol, 1.3 eq.) in ethanol (200 mL) was stirred at reflux for 1-10hours, cooled down to room temperature and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel; hexane/ethyl acetate) giving a target compound (53a-c).Compound 53a: yield 6.89 g, 93% as a yellow solid. APCI-MS (m/z(intensity)): 225.14 ([M+H]⁺, 100%). Compound 53b: yield 7.02 g, 66% asa yellow solid. ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 1.28 (t, 3H), 1.90-1.94(m, 4H), 3.17-3.24 (m, 4H), 4.27 (q, 2H), 7.08 (d, 1H), 7.90 (dd, 1H),8.21 (d, 1H). APCI-MS (m/z (intensity)): 265.16 ([M+H]⁺, 100%). Compound53c: yield 8.80 g, 99% as a yellow solid. ¹H NMR δ_(H) (400 MHz,D₆-DMSO): 1.39 (t, 3H), 3.16 (t, 4H), 3.85 (t, 4H), 4.38 (q, 2H), 7.09(d, 1H), 8.10 (dd, 1H), 8.44 (d, 1H). APCI-MS (m/z (intensity)): 281.14([M+H]⁺, 100%).

Procedure for Synthesis of Compound 53d.

A mixture of 4-chloro-3-nitro-benzoic acid methyl ester (41) (9.92 g,46.00 mmol. 1.0 eq.), phenol (5.20 g, 55.20 mmol, 1.2 eq.), K₂CO₃ (7.63g, 55.20 mmol, 1.2 eq.), CuI (0.26 g, 1.37 mmol, 3 mol %) and DMF (20mL) was stirred at 110° C. for 1.8 hours under argon atmosphere. Thecooled reaction mixture was diluted with water (100 mL). The formedsolid was collected by filtration, dried and purified by columnchromatography (silica gel; hexane/ethyl acetate) giving compound 53d(8.85 g, 70%) as a yellow solid. ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 3.87(s, 3H), 6.98 (d, 1H), 7.09-7.13 (m, 2H), 7.24-7.29 (m, 1H), 7.40-7.46(m, 2H), 8.11 (dd, 1H), 8.59 (d, 1H). APCI-MS (m/z (intensity)): 273.10([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 53e,f.

A mixture of 4-fluoro-3-nitro-benzoic acid ethyl ester (52b) (6.39 g,30.0 mmol, 1.0 eq.), the corresponding amine (30.0 mmol, 1.0 eq.), DIPEA(7.8 mL, 81.6 mmol, 2.7 eq.) and MeCN (60 mL) was stirred at reflux for6-22 hours, cooled down to room temperature and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel; hexane/ethyl acetate) giving a target compound (53e,f).Compound 53e: yield 7.43 g, 90% as a yellow solid. ¹H NMR δ_(H) (400MHz, D₆-DMSO): 1.35 (t, 3H), 2.11 (s, 3H), 4.41 (q, 2H), 6.93 (d, 1H),7.21 (d, 1H), 7.83 (d, 1H), 8.35 (dd, 1H), 8.58 (d, 1H). APCI-MS (m/z(intensity)): 275.69 ([M+H]⁺, 100%). Compound 53f: yield 6.40 g, 86% asan orange solid. ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 1.36 (t, 3H), 4.40 (q,2H), 8.05 (d, 1H), 8.28 (s, 1H), 8.37 (dd, 1H), 8.53 (d, 1H), 9.17 (s,1H). APCI-MS (m/z (intensity)): 263.10 ([M+H]⁺, 100%), 304.17([M+MeCN+H]⁺, 20%).

Generic Procedure for Synthesis of Compounds 53g-i.

To a mixture of 4-chloro-3-nitro-benzoic acid methyl ester (41) (6.26 g,29.07 mmol, 1.0 eq.) and the corresponding boronic acid (R3-B(OH)₂)(43.60 mmol, 1.5 eq.) in toluene (80 mL) and EtOH (80 mL) a mixture ofPd[PPh₃]₄ (725 mg, 0.63 mmol, 0.2 eq.) and an aqueous solution of Na₂CO₃(2M, 35 mL, 70.00 mmol, 2.4 eq.) in toluene (60 mL) and EtOH (60 mL) wasadded rapidly. Then water (35 mL) was added. The reaction mixture wasstirred at reflux under argon atmosphere for 1 hour, then an additionportion of Pd[PPh₃]₄ (725 mg, 0.63 mmol, 0.2 eq.) was added. Theresulted mixture was stirred at reflux for 1-3 hours, cooled down toroom temperature and concentrated at reduced pressure. The obtainedresidue was diluted with water and DCM and filtered through Celite. Theorganic phase was dried over sodium sulfate and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, hexane/ethyl acetate) giving a target compound (53g-i).Compound 53g: yield 3.37 g, 43% as a yellow solid. APCI-MS (m/z(intensity)): 273.14 ([M+H]⁺, 100%), 314.22 ([M+MeCN+H]⁺, 21%). Compound53h: yield 2.44 g, 34% as a yellow solid. APCI-MS (m/z (intensity)):273.13 ([M+H]⁺, 100%). Compound 53i: yield 1.89 g, 70% as a yellowsolid. APCI-MS (m/z (intensity)): 275.75 ([M+H]⁺, 100%), 317.20([M+MeCN+H]⁺, 16%).

Generic Procedure for Synthesis of Compounds 54a-i.

A mixture of the corresponding nitro derivative (53a-i) (29.21 mmol, 1.0eq.), Raney nickel catalyst (3.00 g, 51.12 mmol, 1.75 eq.) and EtOH (450mL).) was stirred under hydrogen atmosphere (1-20 atm.) at roomtemperature for 8-70 hours. The catalyst was removed by filtration. Thefiltrate was concentrated at reduced pressure. The obtained residue waspurified by column chromatography (silica gel, hexane/ethyl acetate)giving a target compound (54a-i). Compound Ma: yield 3.37 g, 59% as awhite solid. APCI-MS (m/z (intensity)): 194.80 ([M+H]⁺, 100%). Compound54b: yield 2.86 g, 51% as a brownish solid. ¹H NMR δ_(H) (400 MHz,CDCl₃): 1.30 (t, 3H), 1.82-1.95 (m, 4H), 3.13-3.20 (m, 4H), 3.72 (brs,2H), 4.30 (q, 2H), 6.87 (d, 1H), 7.24 (s, 1H), 7.41 (d, 1H). APCI-MS(m/z (intensity)): 235.23 ([M+H]⁺, 100%). Compound 54c: yield 3.59 g,46% as a white solid. APCI-MS (m/z (intensity)): 257.17 ([M+H]⁺, 100%).¹H NMR δ_(H) (400 MHz, D₆-DMSO): 1.35 (t, 3H), 2.95 (t, 4H), 3.87 (t,4H), 3.96 (brs, 2H), 4.32 (q, 2H), 6.96 (d, 1H), 7.25 (s, 1H), 7.41 (s,1H), 7.47 (d, 1H). Compound 54d: yield 7.03 g, 93% as a white solid. ¹HNMR δ_(H) (400 MHz, CDCl₃): 3.83 (s, 3H), 3.94 (brs, 2H), 6.80 (d, 1H),7.00-7.05 (m, 2H), 7.10-7.16 (m, 1H), 7.327.40 (m, 3H), 7.51 (d, 1H).APCI-MS (m/z (intensity)): 243.71 ([M+H]⁺, 100%), 285.17 ([M+MeCN+H]⁺,53%). Compound 54e: yield 6.18 g, 93% as a yellowish solid. ¹H NMR δ_(H)(400 MHz, CDCl₃): 1.37 (t, 3H), 2.22 (s, 3H), 3.71 (brs, 2H), 4.38 (q,2H), 6.92 (s, 1H), 7.12 (d, 1H), 7.24 (s, 1H), 7.48 (dd, 1H), 7.52 (d,1H). APCI-MS (m/z (intensity)): 246.14 ([M+H]⁺, 100%). Compound 54f:yield 3.91 g, 72% as a yellowish solid. APCI-MS (m/z (intensity)):233.14 ([M+H]⁺, 100%), 274.19 ([M+MeCN+H]⁺, 19%). Compound Mg: yield1.80 g, 60% as a yellow oil. APCI-MS (m/z (intensity)): 243.77 ([M+H]⁺,100%). Compound 54h: yield 1.99 g, 92% as a yellowish solid. APCI-MS(m/z (intensity)): 242.81 ([M+H]⁺, 100%), 284.19 ([M+MeCN+H]⁺, 42%).Compound 54i: yield 1.50 g, 90% as a yellowish solid. APCI-MS (m/z(intensity)): 246.19 ([M+H]⁺, 100%), 286.56 ([M+MeCN+H]⁺, 22%).

Generic Procedure for Synthesis of Compounds 55a-i.

A solution of sodium nitrite (0.895 g, 12.97 mmol, 1.0 eq.) in water (13mL) was slowly added to a stirred suspension of the corresponding aminederivative (54a-i) (12.78 mmol, 1.0 eq.) in a mixture of concentratedaqueous HCl solution (13 mL) and water (13 mL) at 0° C. By the end ofaddition of the sodium nitrite solution the reaction mixture becameclear. After the addition formation of a precipitate was observed. Thereaction mixture was stirred at 3° C. for 10 minutes after the addition.Then a solution of potassium iodide (4.22 g, 25.42 mmol, 2.0 eq.) inwater (13 mL) was added to the mixture at 3° C. A very viscous red-brownmixture was formed which turned to dark brown color. The reactionmixture was stirred at room temperature for 30 minutes, treated withsaturated aqueous potassium carbonate solution to reach pH>8 andextracted with DCM. The organic layer was washed with an aqueous NaHSO₃solution, with water, dried over sodium sulfate and concentrated atreduced pressure. The obtained residue was purified by columnchromatography (silica gel, hexane/ethyl acetate, 10:1) giving a targetcompound (55a-i). Compound 55a: yield 1.30 g, 25% as a yellow oil.APCI-MS (m/z (intensity)): 306.01 (M+H]⁺, 100%). Compound 55b: yield2.78 g, 70% as a yellowish solid. ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 1.27(t, 3H), 1.85-1.91 (m, 4H), 3.41-3.47 (m, 4H), 4.23 (q, 2H), 6.84 (d,1H), 7.76 (dd, 1H), 8.30 (s, 1H). APCI-MS (m/z (intensity)): 346.14([M+H]⁺, 100%). Compound 55c: yield 3.00 g, 65% as a yellow solid.APCI-MS (m/z (intensity)): 348.04 ([M+H]⁺, 100%). Compound 55d: yield4.39 g, 50% as a yellow solid. APCI-MS (m/z (intensity)): 354.15 (82%),396.09 ([M+MeCN+H]⁺, 100%). Compound 55e: yield 1.40 g, 22% as ayellowish solid. ¹H NMR δ_(H) (400 MHz, CDCl₃): 1.43 (t, 3H), 2.20 (s,3H), 4.44 (q, 2H), 6.87 (d, 1H), 7.08 (d, 1H), 7.36 (d, 1H), 8.13 (dd,1H), 8.62 (d, 1H). APCI-MS (m/z (intensity)): 357.07 ([M+H]⁺, 100%).Compound 55f: yield 4.40 g, 77% as a yellow solid. ¹H NMR δ_(H) (400MHz, CDCl₃): 1.43 (t, 3H), 4.43 (q, 2H), 7.48 (d, 1H), 8.13-8.16 (m,2H), 8.48 (s, 1H), 8.65 (d, 1H). APCI-MS (m/z (intensity)): 343.99([M+H]⁺, 100%), 384.97 ([M+MeCN+H]⁺, 85%). Compound 55g: yield 1.48 g,56% as a yellow oil. APCI-MS (m/z (intensity)): 354.08 ([M+H]⁺, 100%),395.10 ([M+MeCN+H]⁺, 23%). Compound 55h: yield 1.71 g, 59% as ayellowish solid. APCI-MS (m/z (intensity)): 354.00 ([M+H]⁺, 100%),394.96 ([M+MeCN+H]⁺, 39%). Compound 55i: yield 1.76 g, 81% as a yellowoil. APCI-MS (m/z (intensity)): 357.05 ([M+H]⁺, 100%), 398.12([M+MeCN+H]⁺, 48%).

Generic Procedure for Synthesis of Compounds 56a-e,g,i,j.

Triethylamine (5 mL), t-Bu₃P (200 mg, 1.00 mmol, 10 mol %) andPdCl₂[PPh₃]₂ (202 mg, 0.29 mmol, 3 mol %) were added to a solution ofthe corresponding iodide derivative (55a-h) (8.17 mmol, 1.0 eq.) inanhydrous DMF (10 mL). The resulting mixture was stirred under argonatmosphere at room temperature for 10 minutes. Then phenyl-acetylene(1.25 g, 12.25 mmol, 1.5 eq.) was added. The reaction mixture wasstirred at 80° C. for 1-4.5 hours, cooled down to room temperature,diluted with water (30 mL) and extracted with DCM. The organic phase wasdried over sodium sulfate and concentrated at reduced pressure. Theobtained residue was purified by column chromatography (silica gel,hexane/ethyl acetate) giving a target compound (56a-e,g,i,j). Compound56a: yield 0.98 g, 83% as a yellowish oil. APCI-MS (m/z (intensity)):280.19 ([M+H]⁺, 100%). Compound 56b: yield 1.84 g, 72% as a yellowishsolid. ¹H NMR δ_(H) (400 MHz, CDCl₃): 1.38 (t, 3H), 1.97-2.03 (m, 4H),3.73-3.78 (m, 4H), 4.34 (q, 2H), 6.60 (d, 1H), 7.29-7.37 (m, 3H),7.45-7.48 (m, 2H), 7.82 (dd, 1H), 8.12 (d, 1H). APCI-MS (m/z(intensity)): 320.15 ([M+H]⁺, 100%). Compound 56c: yield 2.44 g, 89% asa yellowish solid. ¹H NMR δ_(H) (400 MHz, CDCl₃: 1.40 (t, 3H), 3.38 (t,4H), 3.92 (t, 4H), 4.37 (q, 2H), 6.92 (d, 1H), 7.33-7.39 (m, 3H),7.48-7.53 (m, 2H), 7.94 (dd, 1H), 8.18 (d, 1H). APCI-MS (m/z(intensity)): 366.30 ([M+H]⁺, 100%). Compound 56d: yield 3.18 g, 78% asa yellowish solid. APCI-MS (m/z (intensity)): 329.25 ([M+H]⁺, 100%),370.29 ([M+MeCN+H]⁺, 80%). Compound 56e: yield 0.33 g, 67% as ayellowish solid. APCI-MS (m/z (intensity)): 331.29 ([M+H]⁺, 100%).Compound 56g: yield 1.55 g, 85% as a yellowish solid. APCI-MS (m/z(intensity)): 318.24 ([M+H]⁺, 100%), 359.26 ([M+MeCN+H]⁺, 13%). Compound56i: yield 1.18 g, 92% as a yellowish solid. APCI-MS (m/z (intensity)):328.21 ([M+H]⁺, 100%), 369.23 ([M+MeCN+H]⁺, 20%). Compound 56j: yield1.00 g, 95% as a yellowish solid. APCI-MS (m/z (intensity)): 328.26([M+H]⁺, 100%).

Generic Procedure for Synthesis of Compounds 56f,h,k,l.

Triethylamine (5 mL), CuI (16 mg, 0.084 mmol, 4 mol %) and PdCl₂[PPh₃]₂(50 mg, 0.071 mmol, 3 mol %) were added to a solution of thecorresponding iodide derivative (55e,f,h,i) (2.00 mmol, 1.0 eq.) in MeCN(20 mL). The resulting mixture was stirred under argon atmosphere atroom temperature for 5 minutes. Then 4-fluorophenyl-acetylene (360 mg,3.00 mmol, 1.5 eq.) was added. The reaction mixture was stirred atrefluxing for 2.5-4 hours, cooled down to room temperature andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, hexane/ethyl acetate) giving a targetcompound (56f,h,k,l). Compound 56f: yield 0.25 g, 72% as a yellowishsolid. APCI-MS (m/z (intensity)): 349.20 ([M+H]⁺, 100%). Compound 56h:yield 0.64 g, 95% as a yellowish solid. APCI-MS (161) (m/z (intensity)):336.20 ([M+H]⁺, 100%), 377.22 ([M+MeCN+H]⁺, 52%). Compound 56k: yield0.49 g, 80% as a yellowish solid. APCI-MS (m/z (intensity)): 346.21([M+H]⁺, 100%). Compound 56l: yield 0.36 g, 70% as a yellowish solid.APCI-MS (m/z (intensity)): 349.24 ([M+H]⁺, 100%), 390.29 ([M+MeCN+H]⁺,15%).

Generic Procedure for Synthesis of Compounds 57a-l.

A solution of NaOH (1.48 g, 37.00 mmol, 5.3 eq.) in water (15 mL) wasadded to a suspension of the corresponding ester (56a-1) (7.24 mmol, 1.0eq.) in EtOH (150 mL). The reaction mixture was stirred at 55-60° C. for30 minutes-5.5 hours, cooled down to room temperature, concentrated atreduced pressure, diluted with water (100 mL) and acidified with anaqueous HCl solution (1M) to reach pH 5. The formed precipitate wascollected by filtration and dried giving a target compound (57a-1).Compound 57a: yield 442 mg, 48% as a yellow solid. Compound 57b: yield314 mg, 60% as a white solid. APCI-MS (m/z (intensity)): 291.80 ([M+H]⁺,100%). Compound 57c: yield 2.00 g, 90% as a white solid. APCI-MS (m/z(intensity)): 307.90 ([M+H]⁺, 100%). Compound 57d: yield 457 mg, 15% asa yellowish solid. APCI-MS (m/z (intensity)): 315.20 ([M+H]⁺, 13%),355.89 ([M+MeCN+H]⁺, 100%). Compound 57e: yield 265 mg, 83% as alight-brownish solid. APCI-MS (m/z (intensity)): 303.23 ([M+H]⁺, 100%).Compound 57f: yield 218 mg, 95% as a yellowish solid. APCI-MS (m/z(intensity)): 321.22 ([M+H]⁺, 100%). Compound 57g: yield 1.30 g, 94% asa yellowish solid. ¹H NMR δ_(H) (400 MHz, D₆-DMSO): 3.22 (brs, 1H),7.41-7.48 (m, 5H), 7.79 (d, 1H), 8.10 (dd, 1H), 8.24 (d, 1H), 8.30 (s,1H), 9.24 (s, 1H). APCI-MS (m/z (intensity)): 290.18 ([M+H]⁺, 100%),331.23 ([M+MeCN+H]⁺, 7%). Compound 57h: yield 524 mg, 89% as a yellowishsolid. ¹H NMR (18f) δ_(H) (400 MHz, D₆-DMSO): 3.2 (brs, 1H), 7.41-7.48(m, 5H), 7.79-7.55 (d, 1H), 8.10 (dd, 1H), 8.24 (d, 1H), 8.30 (s, 1H),9.24 (s, 1H). Compound 57i: yield 1.01 g, 94% as a white solid. APCI-MS(m/z (intensity)): 300.15 ([M+H]⁺, 100%), 341.18 ([M+MeCN+H]⁺, 17%).Compound 57j: yield 880 mg, 96% as a white solid. APCI-MS (m/z(intensity)): 300.18 ([M+H]⁺, 100%), 341.03 ([M+MeCN+H]⁺, 10%). Compound57k: yield 420 mg, 90% as a white solid. APCI-MS (m/z (intensity)):318.17 ([M+H]⁺, 100%). Compound 571: yield 320 mg, 95% as a white solid.APCI-MS (m/z (intensity)): 321.19 ([M+H]⁺, 100%), 362.27 ([M+MeCN+H]⁺,8%).

Generic Procedure for Synthesis of Compounds 58a-g, 59a-k.

TBTU (411 mg, 1.28 mmol, 1.5 eq.), triethylamine (0.26 mL, 1.85 mmol,2.2 eq.) and 3-imidazol-1-yl-propylamine (4) or3-(1H-pyrazol-4-yl)-propylamine (6) (0.94 mmol, 1.1 eq.) were added to asolution of the corresponding acid (57a-1) (258 mg, 0.85 mmol, 1.0 eq.)in DCM (10 mL). The reaction mixture was stirred at room temperature for3-15 hours, then diluted with a saturated aqueous NaHCO₃ solution(equivalent volume), again stirred at room temperature for 1 hour andextracted with DCM. The organic phase was dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel; ethyl acetate/MeOH/NH₄OH) giving atarget compound (58a-g, 59a-k). Compound 58a: yield 210 mg, 69% as ayellowish solid. Compound 58b: yield 320 mg, 80% as a yellowish solid.Compound 58c: yield 230 mg, 79% as a yellowish solid. Compound 58d:yield 250 mg, 80% as a yellowish solid. Compound 58e: yield 315 mg, 90%as a yellowish solid. Compound 58f: yield 147 mg, 43% as a yellowishsolid. Compound 58g: yield 170 mg, 60% as a yellowish solid. Compound59a: yield 125 mg, 41% as a yellowish solid. Compound 59b: yield 65 mg,18% as a yellowish solid. Compound 59c: yield 150 mg, 52% as a yellowishsolid. Compound 59d: yield 130 mg, 42% as a yellowish solid. Compound59e: yield 60 mg, 35% as a yellowish solid. Compound 59f: yield 125 mg,37% as a yellowish solid. Compound 59g: yield 85 mg, 52% as a yellowishsolid. Compound 59h: yield 173 mg, 53% as a yellowish solid. Compound59i: yield 80 mg, 28% as a yellowish solid. Compound 59j: yield 115 mg,48% as a yellowish solid. Compound 59k: yield 110 mg, 43% as a yellowishsolid.

Procedure for Synthesis of Compound 62.

TBTU (920 mg, 2.86 mmol, 1.5 eq.), triethylamine (0.42 mL, 3.00 mmol,1.6 eq.) and 3-imidazol-1-yl-propylamine (4) (263 mg, 2.10 mmol, 1.1eq.) were added to a solution of 3-iodo-4-methyl-benzoic acid (61) (500mg, 1.91 mmol, 1.0 eq.) in DCM (20 mL) and DMF (3 mL). The reactionmixture was stirred at room temperature for 20 hours, then diluted witha saturated aqueous NaHCO₃ solution (15 mL), again stirred at roomtemperature for 1.5 hour and extracted with DCM. The organic phase wasdried over sodium sulfate and concentrated at reduced pressure. Theobtained residue was purified by column chromatography (silica gel;ethyl acetate/MeOH/NH₄OH, 40:2:1) giving compound 62 (635 mg, 90%) as ayellowish oil. ¹H NMR δ_(H) (400 MHz, CDCl₃): 2.12 (m, 2H), 2.45 (s,3H), 3.47(q, 2H), 4.05 (t, 2H), 6.25 (brs, 1H), 6.96 (s, 1H), 7.09 (s,1H), 7.27 (d, 1H), 7.52 (s, 1H), 7.60 (dd, 1H), 8.17 (d, 1H). APCI-MS(m/z (intensity)): 370.19 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 63.

Compound 63 was prepared according to Procedure for synthesis ofcompound 62 using 3-(1H-pyrazol-4-yl)-propylamine dihydrochloride (6)(340 mg, 1.72 mmol, 0.90 eq.) and triethylamine (1.5 mL, 10.67 mmol, 5.6eq.). Yield 300 mg, 47% as a yellowish oil. APCI-MS (m/z (intensity)):370.15 ([M+H]⁺, 100%), 411.17 ([M+MeCN+H]⁺, 20%).

Procedure for Synthesis of Compound 64.

Triethylamine (2 mL), t-Bu₃P (43 mg, 0.21 mmol, 10 mol %) andPdCl₂[PPh₃]₂ (43 mg, 0.06 mmol, 3 mol %) were added to a solution ofcompound 62 (630 mg, 1.71 mmol, 1.0 eq.) in DMF (4 mL). The resultedmixture was stirred under argon atmosphere at room temperature for 10minutes. Then phenyl-acetylene (261 mg, 2.56 mmol, 1.5 eq.) was added.The reaction mixture was stirred at 75-80° C. for 2 hours, cooled downto room temperature, diluted with water (20 mL) and extracted with DCM.The organic phase was dried over sodium sulfate and concentrated atreduced pressure. The obtained residue was purified by columnchromatography (silica gel, ethyl acetate) giving compound 64 (380 mg,65%) as a yellowish solid.

Procedure for Synthesis of Compound 65.

Compound 65 was prepared according to Procedure for synthesis ofcompound 64 using compound 63 (290 mg, 0.78 mmol 1.0 eq.),phenyl-acetylene (120 mg, 1.17 mmol, 1.5 eq.), t-Bu₃P (20 mg, 0.1 mmol,10 mol %), PdCl₂[PPh₃]₂ (20 mg, 0.03 mmol, 3 mol %), triethylamine (1mL) and DMF (3 mL). Yield 130 mg, 48% as a yellow solid.

Procedure for Synthesis of Compound 66.

Compound 66 was prepared in the same way as its ethyl ester analog 55c.

Procedure for Synthesis of Compound 67.

A solution of NaOH (1.09 g, 27.00 mmol, 5.0 eq.) in water (5 mL) wasadded to a suspension of compound 66 (1.88 g, 5.41 mmol, 1.0 eq.) inMeOH (110 mL). The reaction mixture was stirred at 50-55° C. for 2hours, cooled down to room temperature, concentrated at reducedpressure, diluted with water (70 mL) and acidified with an aqueous HClsolution (1M) to reach pH 5. The formed precipitate was collected byfiltration and dried giving compound 67 (1.66 g, 93%) as a lightyellowish solid. APCI-MS (m/z (intensity)): 334.11 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 68.

TBTU (1.93 g, 6.00 mmol, 1.5 eq.), triethylamine (2.1 mL, 15.00 mmol,3.7 eq.), 3-(1H-pyrazol-4-yl)-propylamine dihydrochloride (6) (1.03 g,5.20 mmol, 1.3 eq.) were added to a solution of compound 67 (1.33 g,4.00 mmol, 1.0 eq.) in DCM (40 mL). The reaction mixture was stirred atroom temperature for 12 hours, then diluted with a saturated aqueousNaHCO₃ solution (40 mL), again stirred at room temperature for 1.5 hourand extracted with DCM. The organic phase was dried over sodium sulfateand concentrated at reduced pressure. The obtained residue was purifiedby column chromatography (silica gel; ethyl acetate/MeOH/NH₄OH, 40:2:1)giving compound 68 (1.00 g, 57%) as a yellowish oil. APCI-MS (m/z(intensity)): 441.31 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 69.

Triethylamine (0.2 mL, 1.42 mmol, 3.5 eq.), CuI (7 mg, 0.037 mmol, 4 mol%) and PdCl₂[PPh₃]₂ (12 mg, 0.017 mmol, 2 mol %) were added to asolution of compound 68 (686 mg, 2.00 mmol, 1.0 eq.) in MeCN (20 mL).The resulting mixture was stirred under argon atmosphere at roomtemperature for 5 minutes. Then 4-fluorophenyl-acetylene (75 mg, 0.6mmol, 1.5 eq.) was added. The reaction mixture was stirred at refluxingfor 3 hours, cooled down to room temperature and concentrated at reducedpressure. The obtained residue was purified by column chromatography(silica gel, ethyl acetate/MeOH, 10:1) giving compound 69 (108 mg, 62%)as a white solid.

Procedure for Synthesis of Compound 73.

A mixture of 3-bromo-4-imidazol-1-yl-benzoic acid methyl ester (71) (280mg, 1.00 mmol, 1.0 eq.), 4-fluorophenylacetylene (160 mg, 1.33 mmol, 1.3eq.), PdCl₂[PPh₃]₂ (35 mg, 0.05 mmol, 5 mol %), CuI (10 mg, 0.05 mmol, 5mol %) and triethylamine (0.5 mL) in MeCN (7 mL) was refluxed underargon atmosphere for 4 hours. Then an additional amount of PdCl₂[PPh₃]₂(35 mg, 0.05 mmol, 5 mol %), CuI (10 mg, 0.05 mmol, 5 mol %) and4-fluorophenylacetylene (60 mg, 0.50 mmol, 0.5 eq.) were added. Theresulted mixture was refluxed for 4 hours, cooled down to roomtemperature and concentrated at reduced pressure. The obtained residuewas purified by column chromatography (silica gel, DCM/ethyl acetate) togive the intermediate 72 (165 mg) as white solid used on the next stagewithout characterization. To a solution of the intermediate 72 (165 mg,0.52 mmol) in MeOH (20 mL) was added a solution of NaOH (200 mg, 5.00mmol) in water (10 mL). The reaction mixture was stirred at 50° C. for 1hour, cooled down to room temperature, acidified with concentratedaqueous HCl solution to reach pH 4-5. The formed precipitate wascollected by filtration, washed with cold water and diethyl ether anddried to give compound 73 (90 mg, 30% for 2 steps) as beige solid.APCI-MS (m/z (intensity)): 307.12 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 74.

A mixture of compound 73 (90 mg, 0.29 mmol, 1.0 eq.), TBTU (113 mg, 0.35mmol, 1.2 eq.), triethylamine (0.21 mL, 1.45 mmol, 5.0 eq.) was stirredat room temperature in dry DMF (5 mL) for 5 minutes. Then3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (57 mg, 0.35 mmol, 1.2eq.) was added. The resulted mixture was stirred at room temperature for8 hours, diluted with water (100 mL) and extracted with ethyl acetate(3×50 mL). The organic layers were combined, washed with an aqueousK₂CO₃ solution (30 mL), water (3×30 mL), dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, DCM/MeOH) to give compound 74 (72 mg,60%) as a white solid.

Procedure for Synthesis of Compound 77.

A mixture of 3-phenylethynyl-4-(3,4,5-trimethyl-pyrazol-1-yl)-benzoicacid (76) (100 mg, 0.30 mmol, 1.0 eq.), TBTU (126 mg, 0.39 mmol, 1.3eq.), triethylamine (0.23 mL, 1.65 mmol, 5.5 eq.) in dry DMF (5 mL) wasstirred at room temperature for 5 minutes. Then3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (63 mg, 0.39 mmol, 1.3eq.) was added. The resulted mixture was stirred at room temperature for12 hours, diluted with water (100 mL), extracted with ethyl acetate(3×50 mL). The organic layers were combined, washed with an aqueousK₂CO₃ solution (30 mL), water (3×30 mL), dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, DCM/MeOH) to give compound 77 (85 mg,65%) as a beige solid.

Procedure for Synthesis of Compound 79.

A mixture of 3-iodo-4-imidazol-1-yl-benzoic acid methyl ester (71) (492mg, 1.50 mmol, 1.0 eq.), 4-pyridylacetylene (206 mg, 2.00 mmol, 1.3eq.), PdCl₂[PPh₃]₂ (53 mg, 0.075 mmol, 5 mol %), CuI (14 mg, 0.075 mmol,5 mol %), triethylamine (0.5 mL) and MeCN (10 mL) was refluxed underargon atmosphere for 6 hours, cooled down to room temperature andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, DCM/EtOH, 40:1) to give compound 79(430 mg, 94%) as a brownish-grey solid. APCI-MS (m/z (intensity)):304.07 ([M+H]⁺, 100%).

Procedure for Synthesis of Compound 80.

To a solution of compound 79 (430 mg, 1.42 mmol) in MeOH (30 mL) wasadded a solution of NaOH (200 mg, 5.00 mmol) in water (10 mL). Thereaction mixture was stirred at 50° C. for 1 hour, cooled down to roomtemperature, acidified with concentrated aqueous HCl solution to reachpH 4-5. The formed precipitate was collected by filtration, washed withcold water and diethyl ether and dried to give compound 80 (349 mg, 86%)as a grey solid. APCI-MS (m/z (intensity)): 290.09 ([M+H]⁺, 100%).APCI-MS (m/z (intensity)): 288.04 ([M−H]⁻, 100%).

Procedure for Synthesis of Compound 81.

Compound 81 was prepared according to Procedure for synthesis ofcompound 74 using compound 80 (150 mg, 0.52 mmol, 1.0 eq.), TBTU (200mg, 0.62 mmol, 1.2 eq.), triethylamine (0.36 mL, 2.60 mmol, 5.0 eq.) and3-(1H-pyrazol-4-yl)-propylamine hydrochloride (6) (100 mg, 0.62 mmol,1.2 eq.). Yield 91 mg, 44% as a white solid.

TABLE 13 Product com- Yield R- pound ASE code TH code (%) 1 4-Py— 83aASE TH-03631-1 57 51126871 2

83b ASE 51130785 TH-03655-1 36 3

83c ASE 51126873 TH-03633-1 35

Procedure for Synthesis of Compounds 83a-c.

Compounds 83a-c were prepared according to Procedure for synthesis ofcompound 69 using the corresponding acetylenes. Compound 83a: yield 85mg, 57% as a brownish solid. Compound 83b: yield 65 mg, 36% as abrownish solid. Compound 83c: yield 60 mg, 35% as a brownish solid.

Procedure for Synthesis of Compound 88.

A mixture of compound 86 (128 mg, 0.44 mmol, 1.00 eq.),4-imidazol-1-ylmethyl-phenylamine (87) (92 mg, 0.53 mmol, 1.2 eq.), TBTU(241 mg, 0.75 mmol, 1.7 eq.), triethylamine (0.2 mL, 1.44 mmol, 3.3eq.), DCM (3 mL) and THF (5 mL) was stirred at room temperature for 4hours, diluted with equal volume of saturated aqueous NaHCO₃ solutionand stirred at room temperature for 2 hours. The resulting mixture wasextracted with DCM. The organic phase was dried over sodium sulfate andconcentrated at reduced pressure. The obtained residue was purified bycolumn chromatography (silica gel, ethyl acetate/MeOH, 20:1) givingcompound 88 (80 mg, 41%) as a yellowish solid.

To a solution of compound 7c (173 mg, 0.42 mmol, 1.0 eq.) in dry DMF (2mL) were added K₂CO₃ (87 mg, 0.63 mmol, 1.5 eq.) and MeI (239 mg, 1.68mmol, 4.0 eq.). The mixture was stirred at 40° C. for 24 hours, dilutedwith water (100 mL) and extracted with ethyl acetate (3×50 mL). Theorganic layers were combined, washed with water (3×30 mL), dried oversodium sulfate and concentrated at reduced pressure. The obtainedresidue was purified by column chromatography (silica gel, DCM/ethylacetate) to give compound 89 (55 mg, 31%) as a yellowish solid.

Experimental part: General experimental methods. LCMS. The LC/MSanalysis was done at Surveyor MSQ (Thermo Fisher Scientific) with APCIionization. 1. Type of HPLC column: Phenomenex Onyx Monolithic C18;25×4.6 mm; Part No: CHO-7645. 2. Solvent for samples dissolution: 50%DMSO, 50% acetonitrile. 3. Flow rate: 1.5 mL/min; column temperature 25°C. 4. Mobile phase: A=0.1% solution of formic acid in water, B=0.1%solution of formic acid in acetonitrile. 5. Gradient:

time, min. A % B % 0.0 100 0 0.1 100 0 2.1 5 95 2.5 5 95 2.6 100 0 4.0100 06. Detection: diode array (PDA), 200-800 nm; photodiode array detector.Detection was carried out in the full ultraviolet-visible range from 200to 800 nm. APCI (+ or/and − ions)—atmospheric pressure chemicalionization ELSD (PL-ELS 2100). 7. Total run time of the method: 4.5 min.8. Injection volume: 2 μL.

NMR: The ¹H NMR spectra were recorded on a MERCURY plus 400 MHzspectrometer (Varian). Chemical shift values are given in ppm relativeto tetramethylsilane (TMS), with the residual solvent proton resonanceas internal standard.

HPLC: The HPLC analysis was done at Agilent 1100 instrument. 1. Type ofHPLC column: Onyx Monolithic C18, 100×4.6 mm. 2. Flow rate: 1 mL/min;column temperature—ambient. 3. Mobile phase: A=0.1% TFA in water, B=0.1%TFA in acetonitrile.

List of abbreviations: Ac—acetyl, MeCO, APCI—atmospheric-pressurechemical ionization, aq.—aqueous, Ar—aryl or argon, atm—atmosphere(s),brs—broad singlet, Bu—butyl, conc.—concentrated, d—doublet,DABCO—1,4-diazabicyclo[2.2.2]octane, DCM—dichloromethane, dd—doublet ofdoublets, DIPEA—diisopropylethylamine, DMF—dimethylformamide,DMSO—dimethylsulfoxide, dppf—1,1′-bis(diphenylphosphino)ferrocene,ELSD—evaporative light scattering detector, Et—ethyl, eq.—equivalent,h—hour(s), HPLC—high-performance liquid chromatography, i-—iso-,i-Pr—i-propyl, m—multiplet, Me—methyl, MeCN—acetonitrile, MHz—megahertz,n-—normal-, n-Bu—n-butyl, min—minute(s), MS—mass-spectrometry,MWI—microwave irradiation, NBS—N-bromosuccinimide, NMR—Nuclear magneticresonance, PDA—photodiode array, Ph—phenyl, Pr—propyl, q—quartet,Ra—Ni—Raney-nickel, RT—room temperature, s—singlet, t—triplet, t-—tert-,TBTU—N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uroniumtetrafluoroborate, t-Bu—tert-butyl, THF—tetrahydrofuran, TMS(tms)—trimethylsilyl, UV—ultraviolet.

Example 3

This example shows a table of compounds of the present disclosure andtheir activity towards two different cell lines (i.e., MV4-11 andU-397). The IC50 is divided into 5 categories: A<1 μM, B 1-5 μM, C 5-10μM, D 10-20 μM, and E>20 μM.

Compound MV4-11|IC50, U-937|IC50, ID uM uM TT-01901 A A TT-01902 D CTT-02683 A A TT-02684 A A TT-02686 C D TT-02689 B B TT-02690 C DTT-02691 D C TT-02692 E A TT-02694 E A TT-02695 A A TT-02707 E ATT-02709 E A TT-02713 E A TT-02715 A A TT-02717 A A TT-02721 E ETT-02731 A A TT-02732 A A TT-02741 A A TT-02745 A A TT-02746 A ATT-02747 D C TT-02749 A A TT-02750 A A TT-02751 A A TT-02752 C DTT-02760 D D TT-02793 E C TT-02796 D D TT-02797 E E TT-02800 D BTT-02801 D B TT-02802 E B TT-02803 E A TT-02804 E A TT-02805 B ATT-02927 D C TT-02928 D C TT-02929 E E TT-02930 E E TT-02931 E DTT-02932 E E TT-02933 E E TT-02935 D C TT-02936 E E TT-02937 E ETT-02938 E E TT-02939 E C TT-02940 E D TT-02941 D C TT-02942 E CTT-02943 D B TT-02944 E D TT-02945 E E TT-02946 E C TT-02947 E ATT-02948 E E TT-02949 D C TT-03071 B B TT-03073 E E TT-03196 A ATT-03197 A A TT-03198 A A TT-03201 A A TT-03203 A A TT-03211 A ATT-03217 A A TT-03221 A A TT-03225 A A TT-03230 A A TT-03232 A ATT-03233 A A TT-03237 E E TT-03242 A A TT-03245 A A TT-03246 A ATT-03248 A A TT-03252 A A TT-03256 A A TT-03261 A A TT-03264 A ATT-03303 A A TT-03304 A A TT-03305 A A TT-03306 A A TT-03308 A ATT-03309 A A TT-03311 A A TT-03312 A A TT-03321 A A TT-03322 A ATT-03323 A A TT-03324 B B TT-03326 A A TT-03327 A A TT-03328 A ATT-03330 A A TT-03331 A A TT-03332 A A TT-03334 A A TT-03337 D ETT-03346 C E TT-03351 C C TT-03354 A A TT-03355 A A TT-03357 A ATT-03359 A A TT-03364 B B TT-03569 A A TT-03569 A A TT-03574 C CTT-03582 A A TT-03585 A A TT-03586 A A TT-03587 A A TT-03588 A ATT-03588 A A TT-03589 A A TT-03590 A A TT-03591 A A TT-03592 A ATT-03594 A A TT-03595 A A TT-03596 A A TT-03596 A A TT-03597 A ATT-03598 A A TT-03599 A A TT-03602 A A TT-03611 A A TT-03620 A ATT-03623 A A TT-03625 A A TT-03626 A A TT-03627 A A TT-03630 A ATT-03631 A A TT-03633 A A TT-03634 A A TT-03655 A A TT-03669 A ATT-03670 A A TT-03671 A A TT-03676 A A TT-03717 A A TT-03718 A ATT-03720 A A TT-03725 A A TT-03727 A A TT-03732 A B TT-03733 A BTT-03749 B D TT-03750 A A TT-03751 A A TT-03752 A A TT-03753 A ATT-03754 A A TT-03756 A TT-03761 A TT-03762 A TT-03765 A A TT-03767 ATT-03768 A TT-03772 A TT-03773 A A TT-03774 A B TT-03782 A A TT-03783 AA

Example 4

This example provides data related to the efficacy of the instantcompounds in xenograft models of human leukemia cells.

Efficacy of compounds was tested in two types of xenograft models ofleukemia: subcutaneous (SC) (development of tumors after SC inoculationof cells) and systemic and disseminated (development of tumors ondifferent organs after intravenous inoculation of cells). MV4-11 cells(ATCC CRL-9591) of acute myelomonocytic leukemia (AML) were used.

Treatment of SCID mice inoculated SC with MV4-11 cells with compoundsTT-03197 and TT-03203 resulted in a dose-dependent reduction of tumorgrowth (FIGS. 1 and 2). The maximum suppression of tumor growth(calculated using formula: STG %=(volume control-volume treated)/volumecontrol*100) was 28% for mice treated with 10 mg/kg of TT-03197 and 55%for mice treated with 40 mg/kg of TT-03203 (FIG. 1). For TT-03203 themaximum STG was 27% for mice treated PO with 10 mg/kg of TT-03203 and46% for mice treated with 25 mg/kg of TT-03203 (FIG. 2).

Systemic (disseminated) leukemia model was shown to be a more accuraterepresentation of human disease. In this model leukemia cells eitherfrom patient of from cell lines are administered intravenously into miceand leukemia cells are engrafted into blood and blood-forming organs(bone marrow, thymus, spleen).

In systemic model of AML SCID mice were irradiated with 3 Gy andinoculated intravenously with MV4-11 cells 24 h after irradiation. Micewere treated with vehicle control, TT-01901 and TT-03586 during days4-58 (FIG. 3). 30 days after cells implantation mice treated withvehicle were gradually becoming sick, scruffy and were losing weight.Necropsy performed on sick mice revealed multiple tumors on internalorgans, enlargement of spleen and liver in some mice. Mice treated withcompounds survived significantly longer. The ratio of survival oftreated mice versus survival of control mice in percent (T/C) wascalculated using formula: T/C %=average survival time for treatedmice/average survival time for control mice*100. Increase in thesurvival (IS) was calculated using formula: IS=(average survival time oftreated mice−average survival time of control mice)/average survivaltime of control mice*100. The survival of mice treated with TT-01901 was135% and with TT-03586 148% of that for mice treated with vehicle. Thecorresponding increase in survival was 35% and 48%.

1. A compound having the following structure:

wherein X is a carbon atom or nitrogen atom, Y is a single or triplebond, R¹ is selected from the group consisting of a hydrogen atom, asubstituted or unsubstituted five to eight membered heterocyclic ring,six membered aryl ring, five or six membered heteroaryl ring, C₃ to C₈cycloalkyl group, C₁ to C₆ alkyl group,

R² is selected from the group consisting of a hydrogen atom, asubstituted or unsubstituted five or six membered heteroaryl ring, afive or six membered aryl ring, C₃ to C₆ cycloalkyl group, eight to tenmembered heterocyclic ring system, and

R³ is selected from the group consisting of a substituted orunsubstituted C₂ to C₈ alkylheteroaryl group, C₂ to C₈alkyleneheteroaryl group, C₆ to C₁₀ aryl group, C₂ to C₅ heteroarylgroup, C₇ to C₁₃ alkylaryl group, C₇ to C₁₃ alkylenearyl group, C₂ to C₈alkylhetrocyclyl group, C₂ to C₈ alkylenehetrocyclyl group, C₄ to C₈alkylcycloalkyl group, C₄ to C₈ alkylenecycloalkyl group, or takentogether with R⁴ and the nitrogen atom to which they are attached form afive to seven membered substituted or unsubstituted heterocyclic ring;R⁴ is selected from the group consisting of a hydrogen atom andsubstituted or unsubstituted C₁ to C₆ alkyl group; and R^(a) is asubstituted or unsubstituted C₁ to C₆ alkyl group or C₅ to C₆ arylgroup.
 2. The compound of claim 1, wherein the compound has thefollowing structure:


3. The compound of claim 1, wherein the compound has the followingstructure:


4. The compound of claim 1, wherein the compound has the followingstructure:

wherein R⁵ is a C₂ to C₅ heteroaryl group.
 5. The compound of claim 1,wherein the compound has the following structure:


6. The compound of claim 1, wherein the compound has the followingstructure:


7. The compound of claim 1, wherein the compound has the followingstructure:


8. The compound of claim 1, wherein the compound has the followingstructure:


9. The compound of claim 1, wherein the compound has the followingstructure:


10. The compound of claim 1, wherein R¹ is

wherein Z is N(R⁶)₂ or OR⁶, where each R⁶ is independently a hydrogenatom or substituted or unsubstituted C₁ to C₆ alkyl group.
 11. Thecompound of claim 1, wherein R² is

wherein each R⁶ is independently a hydrogen atom or substituted orunsubstituted C₁ to C₆ alkyl group and n is 1, 2, 3, or
 4. 12. Thecompound of claim 1, wherein the ring formed by R³—N—R⁴ is selected fromthe following structures:


13. The compound of claim 1, wherein R³ is selected from the followingstructures:


14. The compound of claim 1, wherein R⁴ is a hydrogen atom or a methylgroup.
 15. The compound of claim 1, wherein R¹ is a substituted orunsubstituted five to eight membered heterocyclic ring.
 16. The compoundof claim 15, wherein the unsubstituted five to eight memberedheterocyclic ring comprises at least one nitrogen atom.
 17. The compoundof claim 1, wherein the compound is selected from the group consistingof


18. A method of treating cancer in an individual diagnosed with orsuspected of having cancer comprising administering to the individual atherapeutically effective amount of a compound of claim
 1. 19. Themethod of claim 18, wherein the cancer is a hematopoietic cancer. 20.The method of claim 19, wherein the hematopoietic cancer is leukemia.